Families of transmembrane transporters selective for amino acids and their derivatives

Milton H. Saier, Jr1

Department of Biology, University of California at San Diego, La Jolla, CA 92093-0116, USA1

Tel: +1 858 534 4084. Fax: +1 858 534 7108. e-mail: msaier{at}ucsd.edu

Keywords: transmembrane transport, phylogeny, evolution, amino acids, polyamines

The information presented in this review was initially prepared for presentation at the FASEB meeting on amino acid transport held in Copper Mountain, Colorado, June 26–July 1, 1999 and was updated in January 2000 following the meeting of the Transport Nomenclature Panel of the International Union of Biochemistry and Molecular Biology (IUBMB) in Geneva, November 28–30, 1999. The system of classification described in this review reflects the recommendations of that panel.


   Overview
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Twenty-one families of secondary [proton-motive force (PMF)-driven] carriers are herein described, which, in addition to 13 of the currently recognized 48 families within the ATP-dependent ATP-binding casette (ABC) superfamily and three families of channel proteins, largely account for the transport of amino acids and their derivatives (excluding proteins) into and out of all living cells on Earth. Family characteristics as well as structural and functional properties of the family constituents are described. By reference to our web site (http: / / www-biology.ucsd.edu /~msaier / transport /), sequences, phylogenetic relationships, detailed substrate-specificity information and the distribution of members of each family in 20 fully sequenced genomes of bacteria, archaea and eukaryotes are available. This review provides a comprehensive guide to the diversity of carriers that mediate the transport of amino acids and their derivatives across cell membranes.


   Background
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Amino acids are the building blocks of proteins. They also provide raw materials for a variety of important cellular processes such as energy generation, nitrogen metabolism, cell-wall synthesis, antibiotic production and intercellular communication. Amino acids can be zwitterionic, anionic or cationic, and their side chains can be hydrophobic, semipolar or strongly hydrophilic. These side chains may be aliphatic or aromatic, and they sometimes serve as physiologically important proton donors and acceptors in enzymes with pKa values ranging from about 4 to 12. Amino acids in proteins can be derivatized by methylation, acetylation, formylation and phosphorylation; the free compounds can be decarboxylated to amines, and L-aminoacyl derivatives can be condensed enzymically with other L-amino acids, with amino acids of the D configuration and with hydroxy acids to yield peptides and depsipeptides of unusual composition. These aminoacyl residue derivatives serve a wide variety of biological functions including those of biological warfare, quorum sensing, intercellular signalling and regulation of gene expression. It is therefore not surprising that several transporter families have evolved to control and facilitate exchange of these substances between cells and their environment, between cells and their organelles, and between cells and other living entities.

Over two dozen transporter families and superfamilies are currently recognized that include members that transport amino acids and their derivatives (Paulsen et al., 1998a , b ; Saier, 1998 , 1999a , b , c ; see also http://www-biology.ucsd.edu/~msaier/transport/). A number of interesting questions arise related to these families. For example (1) what are the ranges of substrates transported by members of each of these families? (2) What energy-coupling mechanisms do they employ to accumulate or expel their substrates within or from living cells? (3) What is/are the polarity/polarities of transport allowed by members of each such family? (4) In what range of organisms are members of each of these families found? (5) When, during evolutionary history, did each family appear and via what route? (6) What are the mechanisms used to effect transmembrane transport? (7) Do homologues of these transport proteins function in capacities other than in transport, and if so, in what capacities? (8) Are certain families concerned primarily with a particular physiological function (e.g. uptake, secretion or intercellular communication), or do they all serve comparable and broadly overlapping physiological functions? (9) How are the syntheses, activities, intracellular localizations and proteolytic degradation processes of transporter proteins regulated, and in response to what stimuli? (10) What transport characteristics are readily altered during evolution and which characteristics remain relatively immutable? These and other questions will be systematically addressed in the following section, and information that will allow the reader to personally evaluate the answers to these questions, relevant to specific families, will be provided with greater insight in the subsequent sections.


   Characteristics of transporters and their homologues
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Careful examination of the properties of the functionally characterized members of currently recognized transporter families selective for amino acids and their derivatives reveals tentative answers to the ten questions posed above.

(1) The range of substrates transported by an individual member of a family can be narrow or broad, depending on the transporter, and an entire family can often transport a wide variety of structurally related compounds. Seldom, however, do members of a single family catalyse the transport of structurally divergent types of compounds (i.e. sugars versus amino acids versus inorganic anions).

(2) Only two forms of energy are generally used for the active uptake or expulsion of amino acids and their derivatives: electrochemical energy stored in the gradients of H+, Na+ and structurally related substrates (utilized by secondary active carriers), and chemical energy in the form of ATP (utilized by ABC-type uptake and efflux systems as well as by several macromolecular transport systems). Channel proteins usually do not couple transmembrane translocation to energy, although a few interesting exceptions are known.

(3) With very few exceptions (i.e. the major-facilitator superfamily and the ABC superfamily) all members of a phylogenetically defined family function with strongly preferential inwardly directed or outwardly directed polarity. Seldom do different members of a single transporter family transport solutes with opposite polarity, and no permease is currently known to actively take up its substrates under one set of experimental conditions but actively extrude them under another set of conditions.

(4) Some families of sequence-related transport proteins are found ubiquitously in all major groups of organisms (i.e. among the bacteria, archaea and eukaryotes), while others are restricted to just one of these three domains. In some cases, all members of a family derive from just one kingdom within one of these domains. The former proteins may represent ancient families of transporters while the latter proteins are those which have more recently appeared, either de novo or by extensive sequence divergence from a pre-existing family.

(5) As noted above and discussed in several reviews (Saier, 1994 , 1996 , 1998 ), it appears that new families have appeared throughout evolutionary history although with variable frequencies. Moreover, they probably underwent sequence divergence at rates that were neither constant with time nor constant for all members of the family. Most of the families that appeared after bacteria, archaea and eukaryotes separated from each other during the ‘great split’ are apparently still restricted to the domain in which they arose. These proteins have often arisen from much simpler peptides or proteins as a result of internal gene-duplication and gene-fusion events. Little lateral transfer of the resultant genetic information between the three domains of organisms has occurred, at least during the past two billion years.

(6) The primary modes of transport have historically been thought to involve, and are still considered to involve, channels and carriers. While the mechanistic details of the former type of transporters are well understood, in part due to the availability of high-resolution three-dimensional structural data for these proteins, carrier mechanisms are still poorly understood (West, 1997 ). These carrier mechanisms represent the primary types responsible for the transmembrane translocation of organic nutrients.

(7) Homologues of transporters almost never function in a primary capacity other than transport. Those that do usually serve as simple receptors, influencing a single aspect of cellular physiology. We have consequently proposed that integral transmembrane transporters evolved as a distinct class of proteins, and that they did not appear by modification of other protein types (i.e. enzymes, structural proteins or regulatory proteins) (Saier & Tseng, 1999 ).

(8) Some families or subfamilies of transporters selective for amino acids and their derivatives seem to have evolved to serve a highly specific physiological function such as that of auxin secretion for the purpose of promoting gravitropism in plants, performed by members of the auxin efflux carrier (AEC) family [transporter classification (TC) no. 2.A.69], or that of promoting dormant spore germination in Bacillus species, performed by members of the spore germination protein (SGP) family (TC no. 2.A.3.9) of the amino acid/polyamine/organocation (APC) superfamily (TC no. 2.A.3). Other families selectively catalyse either uptake or efflux of specific amino acids and/or their derivatives for the purpose of nutrition, cellular protection or intercellular signalling. Thus, it is clear that members of a family may have evolved to provide specific physiological functions. Nevertheless, specialization for a specific physiological function appears to be a late-evolving characteristic and therefore is of minimal importance for purposes of classifying transporters (Saier & Tseng, 1999 ).

(9) In general, the syntheses, activities and degradation of transport proteins are stringently regulated, employing the same or similar universal mechanisms (i.e. ligand binding, covalent modification, protein–protein interactions) that are operative and well established for cytoplasmic enzymes. Similarity, subcellular localization is determined by targeting sequences in the proteins of both prokaryotes and eukaryotes, and by complex, reversible, hormone-regulated processes in higher eukaryotes. These topics are in general beyond the scope of this review but are the topics of numerous other reviews, to which the reader is referred (Saier et al., 1989 ; Knutson, 1991 ; Fekkes & Driessen, 1999 ; Koch et al., 1999 ).

(10) Finally, the classification and characterization of transporter families have revealed that transport mode and energy-coupling mechanism are highly conserved evolutionary traits, that protein topology, polarity of transport and substrate specificity are traits conserved to an intermediate degree, and that the regulatory mechanisms imposed on transporters are late evolving, poorly conserved traits. The utilization of these various traits for purposes of transport-protein classification has been the topic of discussion in several recently published reviews (Saier, 1999a , b , c ; Saier & Tseng, 1999 ).

Below, the transporter families concerned with transmembrane transport of amino acids and their derivatives are discussed. Groups of functional types will first be considered according to class and transport mechanism, and the unique features of each such family will be presented.


   Most amino acid transporters are secondary carriers
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
We currently recognize about 120 families of channel/pore proteins, about 30 families of primary active transport systems and about 80 families of secondary carriers (Saier, 1998 , 1999a , b , c ; see also http://www-biology.ucsd.edu/~msaier/transport/). Additionally, dozens of families include transporters with undefined, or ill-defined, transport mechanisms. Of the former families, members of only one recognized family of channel proteins, the phospholemman (PLM) family (TC no. 1.A.27; Kirk & Strange, 1998 ), in addition to nonspecific outer-membrane porins of Gram-negative bacteria and eukaryotic organelles (TC classification division 1.B), and only one recognized family of primary carriers, the ABC superfamily (Saurin et al., 1999 ), can transport amino acids and their structurally related derivatives. All other families apparently consist of secondary carriers that function by PMF or SMF (sodium-ion motive force) -driven uptake, by PMF-driven efflux, by solute–solute exchange or by uniport. Secondary carriers will therefore be the main focus of this review.


   ABC (ATP-binding cassette) -type transporters
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
ABC-type transporters fall into 48 currently recognized families within the ABC superfamily (Table 1). Of these, 19 include members that are exclusively uptake systems (all found in prokaryotes); 19 include members that are prokaryotic-specific efflux systems and 10 include members that are primarily eukaryotic-specific efflux systems. The uptake and efflux systems cluster separately on the phylogenetic tree for this superfamily (Saurin et al., 1999 ).


View this table:
[in this window]
[in a new window]
 
Table 1. The ATP-binding cassette (ABC) superfamily (TC 3.A.1)

 
Of the 19 ABC-type uptake-system families, two are primarily concerned with polar and non-polar amino acid transport (TC 3.A.1.3 and 4, respectively); a third is largely concerned with peptide transport (TC 3.A.1.5); a fourth is primarily concerned with polyamine and opine uptake (TC 3.A.1.11); a fifth is concerned with quaternary amine uptake (TC 3.A.1.12); and a sixth functions to take up taurine (the decarboxylation product of cysteic acid) (TC 3.A.1.17). Many the ABC uptake systems appear to catalyse irreversible vectorial reactions, but one system, the polar amino acid transporter (Aap) of Rhizobium leguminosarum, has been shown to catalyse both active uptake and passive efflux. This system is unusual not only because of its reversibility but also because of its broad specificity (Walshaw & Poole, 1996 ; Walshaw et al., 1997 ). Whether the capacity to catalyse efflux of solutes will prove to be a general characteristic of ABC uptake systems or a specific trait of only a few of these systems is currently an unanswered question requiring further experimental work.

Of the prokaryotic ABC-type exporter families, three are concerned with drug efflux, five catalyse peptide export and two function in protein transport. The five peptide-exporter families are the Pep1–3E families (TC 3.A.1.101–103) and the two microcin-exporter families (the McbE and McjD families; TC 3.A.1.106 and 108 respectively), but some of the drug-export permeases may be capable of transporting certain peptides. Of the currently recognized eukaryotic families, two function primarily in peptide export (a-factor sex pheromone exporter, STE, and MHC peptide transporter, TAP; TC 3.A.1.206 and 209 respectively) and three function to export drugs (including some peptides). However, no eukaryotic ABC exporter is known to transport proteins. Eukaryotes export macromolecules by exocytosis rather than by a permease-mediated molecular-transport process. Thus, the functional diversity of prokaryotic ABC transporters far exceeds that of the eukaryotic ABC transporters. Of the 48 recognized families, 13 (27%) are concerned primarily with the transport of amino acids and their derivatives including peptides, and only two of these families derive from eukaryotes. In bacteria, another two families are concerned with protein export.


   Secondary carriers
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Table 2 tabulates the secondary carrier families that are known to function in transport of (a) amino acids and their conjugates, (b) amines, amides and polyamines and (c) peptides. Altogether, 19 families fall into category a, eight fall into category b, and four fall into category c. Nevertheless, there is a total of only 21 families. All families that fall into category b are also represented in category a and only two families of peptide transporters (oligopeptide transporter, OPT, and peptide-uptake permease, PUP) have not been shown to include members that transport simple amino acids. Below we examine these families in greater detail.


View this table:
[in this window]
[in a new window]
 
Table 2. Families of secondary active transporters specific for amino acids and their derivatives

 

   Secondary amino acid transporter families found exclusively in bacteria
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Table 3 lists and provides characteristics of the ten prokaryotic-specific families of secondary transporters that include members exhibiting specificity for amino acids and their derivatives. These families are almost without exception small, usually with less than 20 currently sequenced members. At least one of these families, the hydroxy and aromatic amino acid porter (HAAAP) family, is probably distantly related to the largest known superfamily of amino acid transporters, the APC superfamily (Young et al., 1999 ; Jack et al., 2000 ).


View this table:
[in this window]
[in a new window]
 
Table 3. Secondary active amino acid transporter families found exclusively in prokaryotes

 
The first two bacterial-specific families listed in Table 3 (C4-dicarboxylate uptake, Dcu, and betaine/carnitine/choline transporter, BCCT) have recently been described in some detail and characterized phylogenetically (Saier et al., 1999a ). Six of the ten families listed in Table 3 exhibit properties that are generally characteristic of secondary solute-uptake transporters: they function by H+ or Na+ symport exclusively in the uptake of their nutrient substrates and they usually consist of single polypeptide chains exhibiting about 12 transmembrane {alpha}-helical spanners (TMSs). In most cases, they function in the absence of auxiliary subunits. One exceptional family is the tripartite ATP-independent periplasmic transporter (TRAP-T) family of dicarboxylate/amino acid uptake permeases (Table 3). TRAP-T family transporters function by a typical H+ symport mechanism, characteristic of secondary carriers, for the uptake of dicarboxylates including the amino acid glutamate. However, in contrast to other secondary carriers, they are normally heterotrimeric with three dissimilar, non-homologous subunits/domains (Forward et al., 1997 ; Rabus et al., 1999 ). In contrast, the characterized proteins of the L-lysine exporter (LysE) and resistance to homoserine and threonine (RhtB) families export amino acids, probably by a proton antiport mechanism (Vrljic et al., 1999 ; Zakataeva et al., 1999 ). These two families are probably constituents of a single superfamily (Vrljie et al., 1999). The available evidence suggests that a single gene product participates in the transport process. Another family, the carboxylate/amino acid/amine transporter (CAAT) family, also includes members that export amino acids and their derivatives. A number of additional prokaryotic-specific families of (putative) transporters included under TC classification category 9 and not presented in Tables 2 and 3 are likely to prove to consist of efflux pumps.

The 10 families listed in Table 3 will be discussed individually below. The descriptions provided reflect our knowledge of these families as of July 1999.

The Dcu (C4-dicarboxylate uptake) family (TC 2.A.13)
Several proteins of the Dcu family have been sequenced, all from Gram-negative bacteria (Engel et al., 1994 ; Six et al., 1994 ; Unden & Bongaerts, 1997 ; Golby et al., 1998 ). The two best characterized systems (DcuA and DcuB) are from Escherichia coli. The fully sequenced proteins of the Dcu family are of fairly uniform size (434–446 residues). They possess 12 putative TMSs, but DcuA of Escherichia coli has 10 experimentally determined TMSs with both the N and C termini localized to the periplasm. For DcuA, the ‘positive inside’ rule (von Heijne, 1992 ) is obeyed and two putative TMSs are localized to a cytoplasmic loop between TMS 5 and 6 and in the C-terminal periplasmic region.

The Escherichia coli DcuA and DcuB proteins transport aspartate, malate, fumarate and succinate, and function as antiporters with any two of these substrates. The two proteins exhibit 36% identity with 63% similarity, and both transport fumarate in exchange for succinate with about the same affinity (30 µM). Since DcuA is encoded near the gene for aspartase, and DcuB is encoded in an operon with the gene for fumarase, their physiological functions have been hypothesized to involve aspartate:fumarate and fumarate:malate exchange during the anaerobic utilization of aspartate and fumarate, respectively. However, the electroneutral antiport of fumarate for succinate during anaerobic fumarate respiration has been demonstrated, and both permeases are induced under anaerobic conditions and subject to catabolite repression. The two transporters can apparently substitute for each other under certain physiological conditions. A third dicarboxylate transporter, DcuC, exhibits slight sequence similarity with DcuA (optimized comparison score of 5 SD; 23% identity for a segment of 100 residues). This degree of similarity is insufficient to establish homology (Saier, 1994 , 1996 ) and consequently, DcuC is assigned to a distinct family (TC 2.A.61).

The BCCT (betaine/carnitine/choline transporter) family (TC 2.A.15)
Proteins of the BCCT family are found in Gram-negative and Gram-positive bacteria (Lamark et al., 1991 ; Eichler et al., 1994 ; Kappes et al., 1996 ; Kempf & Bremer, 1998 ; Peter et al., 1998 ). Their common functional feature is that they all transport molecules with a quaternary ammonium group [R-N+(CH3)3]. BCCT family proteins vary in length between 481 and 677 aminoacyl residues, possess 12 putative TMSs and are energized by PMF-driven proton symport.

The alanine/glycine:cation symporter (AGCS) family (TC 2.A.25)
Members of the AGCS family transport alanine and/or glycine in symport with a monovalent cation, Na+ and/or H+ (see Reizer et al., 1994 ). The proteins are 445–542 aminoacyl residues in length and possess 8–12 putative TMSs. They are found in Gram-positive and Gram-negative bacteria. Only two members of the family have been functionally characterized. These proteins show minimal sequence similarity to members of the APC family (TC 2.A.3; see below; Jack et al. 2000 ), but the significance of this observation is not known.

The branched chain amino acid:cation symporter (LIVCS) family (TC 2.A.26)
Characterized members of this family transport all three of the branched chain aliphatic amino acids [leucine (L), isoleucine (I) and valine (V)] (Reizer et al., 1994 ; Stucky et al., 1995 ; Tauch et al., 1998 ). They are found in Gram-negative and Gram-positive bacteria and function by a Na+ or H+ symport mechanism. They possess about 440 aminoacyl residues and display 12 putative TMSs.

The glutamate:Na+ symporter (GltS) family (TC no. 2.A.27)
A single member of this family has been functionally characterized (Deguchi et al., 1990 ). This permease (GltS of Escherichia coli) catalyses glutamate:Na+ symport. It exhibits 401 aminoacyl residues with 12 putative helical TMSs. Homologues are found in other bacteria including Haemophilus influenzae, Helicobacter pylori and Synechocystis.

The HAAAP (hydroxy and aromatic amino acid porter) family (TC 2.A.42)
The HAAAP family includes three well characterized aromatic amino acid:H+ symport permeases of Escherichia coli: a high-affinity tryptophan-specific permease, Mtr, a low affinity tryptophan permease, TnaB, and a tyrosine-specific permease, TyrP (Wookey & Pittard, 1988 ; Sarsero et al., 1991 ; Sarsero & Pittard, 1995 ). These proteins possess 403–405 aminoacyl residues with 11 TMSs. The HAAAP family also includes two well characterized permeases specific for hydroxy amino acids: one is the serine permease, SdaC, of Escherichia coli, the other is the threonine permease, TdcC, of Escherichia coli (Goss et al., 1988 ; Shao et al., 1994 ). These permeases are 429 and 443 aminoacyl residues long, respectively, and each possesses 11 putative TMSs. Homologues are present in a variety of Gram-negative bacteria. They show topological features common to members of the eukaryotic amino acid/auxin permease (AAAP) family (TC 2.A.18), and they display limited sequence similarity with some of them (Young et al., 1999 ). Since members of the AAAP family show limited sequence similarity with members of the large APC superfamily (TC no. 2.A.3) (Young et al., 1999 ; Jack et al., 2000 ), all of these proteins are probably related.

The TRAP-T (tripartite ATP-independent periplasmic transporter) family (TC 2.A.56)
TRAP-T family permeases generally consist of three components (Forward et al., 1997 ; Rabus et al., 1999 ). The best characterized of these systems is the DctMQP system of Rhodobacter capsulatus (Forward et al., 1997 ). DctM is a typical 12 TMS protein with weak sequence and motif similarity to several other families of secondary carriers (Rabus et al., 1999 ). However, DctQ is a 4 TMS integral membrane protein, and DctP is a periplasmic binding protein. Database searches and phylogenetic analyses have revealed that unlike most of the other families included in Table 3, homologous TRAP-T systems are found in archaea as well as bacteria. In some of these permeases, including all currently recognized archaeal systems, the M homologue is fused to the Q homologue yielding a 16 TMS protein, and in a few cases, the Q homologue is fused to the P homologue. Thus, in the TRAP-T family, as for the ABC superfamily, domain fusion/splicing has occurred repeatedly during evolution of the family.

All three Dct proteins are required for the uptake of dicarboxylates in Rhodobacter capsulatus, suggesting that these subunits function together, employing a concerted mechanism. Moreover, phylogenetic analyses reveal that all TRAP-T systems probably depend on the structural equivalent of the three proteins, DctMQP. The family appears to be an ancient but functionally unified family that evolved in parallel with the ABC superfamily, with little or no shuffling of constituents between families (Rabus et al., 1999 ). The TRAP-T family is the only family currently known for which an extracytoplasmic receptor functions in conjunction with a secondary carrier.

Many members of the TRAP-T family are functionally uncharacterized. An operon encoding a Synechocystis system includes a protein homologous to the glutamine-binding protein of an Escherichia coli ABC-type permease, and biochemical evidence has suggested that a glutamate transporter from Rhodobacter sphaeroides is a periplasmic binding protein-dependent, PMF-dependent, secondary carrier (Jacobs et al., 1996 ). Escherichia coli homologues may be involved in the uptake of pentoses and/or pentitols (Reizer et al., 1996 ; Sanchez et al., 1994 ). Thus, members of the TRAP-T family of permeases may take up widely divergent compounds.

The LysE (L-lysine exporter) family (TC 2.A.75)
One member of the LysE family (LysE of Corynebacterium glutamicum) is functionally well characterized, but functionally uncharacterized or partially characterized homologues are encoded within the genomes of a variety of bacteria and archaea (Bröer & Krämer 1991a , b ; Vrljic et al., 1996 , 1999 ; Zakataeva et al., 1999 ). All of these proteins are 190–240 aminoacyl residues in length and possess six hydrophobic regions. PhoA fusion analyses provided evidence that LysE of Corynebacterium glutamicum exhibits a 5 TMS topology, with the N terminus inside and the C terminus outside (Vrljic et al., 1999 ). LysE appears to catalyse unidirectional efflux of L-lysine and other basic amino acids such as L-arginine and L-ornithine, and it provides the sole route for L-lysine excretion in this bacterium. The energy source is believed to be the PMF (H+ antiport or OH- symport).

The RhtB (resistance to homoserine and threonine) family (TC 2.A.76)
Distant homologues of LysE have been shown to expel threonine. They may catalyse efflux of homoserine and homoserine lactones as well (Aleshin et al., 1999 ; Zakataeva et al., 1999 ). Because homoserine lactones are signalling molecules in Gram-negative bacteria, this observation has special significance (Fuqua et al., 1996 ; Swift et al., 1996 ). The family that includes these amino acid efflux pumps has been termed the RhtB family, and in the transporter-classification system, it was given the TC number 2.A.76. Evidence has been presented that it and the LysE family (above), together with the cadmium-resistance (CadD) family (TC no. 2.A.77), compose a single superfamily, the LysE superfamily (Vrljic et al., 1999 ). All characterized members of this superfamily catalyse solute export, presumably by a proton-antiport mechanism.

The CAAT (carboxylate/amino acid/amine transporter) family (TC 2.A.78)
The CAAT family is a large family of integral membrane proteins with sizes ranging from 287 to 310 aminoacyl residues and exhibiting 10 putative TMSs. These proteins are derived from phylogenetically divergent bacteria and archaea, and Escherichia coli, Bacillus subtilis and Aspergillus fulgidus have multiple paralogues. With one PSI-BLAST iteration, they show low degrees of sequence similarity with members of the ubiquitous L-rhamnose transporter (RhaT) family (TC 2.A.9) and with the eukaryotic triose phosphate transporter (TPT) family (TC 2.A.50), found in plant chloroplasts and plastids as well as in yeast. One distant plant homologue is the Medicago nodulin N21-like protein (gbAC004218; 374 aa) of Arabidopsis thaliana.

Proteins of the CAAT family evidently arose by an internal gene-duplication event as the first halves of these proteins are homologous to the second halves. Although none of these prokaryotic proteins is functionally characterized, several members of the CAAT family have been implicated in solute transport. Thus, the MttP protein of the archaeon Methanosarcina barkeri may transport methylamine (Ferguson & Krzycki, 1997 ) and the YtfF protein of Chlamydia trachomatis may transport basic amino acids (Stephens et al., 1998 ). Additionally, MadN is encoded within the malonate utilization operon of Malonomonas rubra and PecM is encoded within a locus of Erwinia chrysanthemi controlling pectinase, cellulase and blue-pigment production. MadN may be an acetate-efflux pump while PecM might export the pigments produced by gene products encoded in the pecM operon (Berg et al., 1997 ; Reverchon et al., 1994 ).


   Amino acid transporter families that have been found in eukaryotes
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
While Table 3 lists the properties of the ten families of amino acid transporters found exclusively in prokaryotes, Table 4 provides comparable information for the nine families that occur in eukaryotes. Of these families, the AAAP and mitochondrial carrier (MC) families are found only in eukaryotes. The MC family undoubtedly arose in eukaryotes in response to a need for a new type of communication between the matrix of the mitochondrion and the cytoplasm of the eukaryotic cell (Kuan & Saier, 1993 ; Saier, 1994 , 1996 ). However, we have presented evidence (Young et al., 1999 ) that the AAAP family is distantly related to the ubiquitous APC superfamily, and this claim has recently been further substantiated (Jack et al., 2000 ).


View this table:
[in this window]
[in a new window]
 
Table 4. Secondary active amino acid transporter families found in eukaryotes

 
The AEC family is of particular physiological interest. It is apparently a ubiquitous family found in bacteria, archaea and eukaryotes, but only its members from plants have been characterized. These plant proteins serve a single function: to catalyse polarized auxin efflux for the purpose of promoting gravitropism. This therefore represents an example where a specific subfamily of a larger family has become specialized for a particular physiological function.

Below, the nine families described in Table 4 are discussed.

The APC (amino acid/polyamine/organocation) family (TC 2.A.3)
The APC family of transport proteins includes nearly 250 currently sequenced members that function as solute:cation symporters and solute:solute antiporters (Closs et al., 1993 ; Reizer et al., 1993a ; Sophianopoulou & Diallinas, 1995 ; Isnard et al., 1996 ; Kashiwagi et al., 1997 ; Brechtel & King, 1998 ; Hu & King, 1998a , b ; Sanders et al., 1998 ; Sato et al., 1999 ). All functionally characterized members thus catalyse active solute uptake or exchange transport. These proteins occur in bacteria, archaea, yeast, fungi, unicellular eukaryotic protists, plants and animals, and are thus essentially ubiquitous. They vary dramatically in length, being as small as 350 residues and as large as 800 residues. The smaller proteins are generally of prokaryotic origin while the larger ones are of eukaryotic origin. Most of them possess 12 TMSs, but predicted topologies of 10 or 14 TMSs are also sometimes observed (Cosgriff & Pittard, 1997 ; Hu & King, 1998c ; Jack et al., 2000 ). The larger eukaryotic proteins have N- and C-terminal extensions that may serve regulatory functions. At least some animal proteins such as ASUR4 (gbY12716) and SPRM1 (gbL25068) associate with a type 1 transmembrane glycoprotein that is essential for insertion, stability or activity of the permease and forms a disulfide bridge with it (Verrey et al., 1999 ). These glycoproteins include the CD98 heavy-chain protein of Mus musculus (gbU25708) and the 4F2 cell surface antigen heavy chain of man (Mastroberardino et al., 1998 ). These homologues are members of the rBAT family of mammalian transporter-accessory proteins (TC 8.A.9). All of the members of the rBAT family are believed to associate with members of the L-type amino acid transporter (LAT) family (TC 2.A.3.8) within the APC superfamily to form heterodimeric complexes.

One APC family member, Hip1 of Saccharomyces cerevisiae, which clearly functions in amino acid transport, has also been implicated in heavy metal transport (Farcasanu et al., 1998 ). Another member of the family, Ssy1 of Saccharomyces cerevisiae, appears to be a transcriptional regulatory sensor (Didion et al., 1998 ). It is possible, but not demonstrated that APC family homologues in Bacillus subtilis that promote spore germination may function as receptors rather than transporters. Thus, a few APC family members may serve functions other than that of the transport of amino acids and their derivatives.

The AAAP (amino acid/auxin permease) family (TC 2.A.18)
The AAAP family includes over four dozen sequenced proteins from plants, animals, yeast and fungi (Fischer et al., 1995 ; Bennett et al., 1996 ; Rentsch et al., 1996 ; McIntire et al., 1997 ; Young et al., 1999 ). Individual permeases of the AAAP family transport auxin (indole-3-acetic acid), {gamma}-aminobutyric acid, a single L-amino acid or multiple amino acids. Some of these permeases exhibit very broad specificities, transporting all of the L-amino acids naturally found in proteins. There are 16 AAAP paralogues in Saccharomyces cerevisiae, 13 in Caenorabditis elegans and at least 9 in Arabidopsis thaliana. These proteins, all from eukaryotes, vary from 376 to 713 aminoacyl residues in length, but most are 400–500 residues. Most of the size variation occurs as a result of the presence of long N-terminal hydrophilic extensions in some of the proteins, but size variation in the loops and the C termini is sometimes observed. These proteins exhibit 11 putative TMSs and show limited sequence similarity with members of the large APC superfamily. Thus, the AAAP family is probably part of the APC superfamily (Jack et al., 2000 ).

The major facilitator superfamily (MFS) (TC 2.A.1)
The MFS is a very old, large and diverse superfamily that includes over a thousand sequenced members (Pao et al., 1998 ; Saier et al., 1999b ). These permeases catalyse uniport, solute:cation (H+ or Na+) symport and/or solute:H+ or solute:solute antiport. Most are of 400–600 aminoacyl residues in length and possess either 12 or 14 putative or established TMSs. They exhibit specificity for sugars, polyols, drugs, neurotransmitters, Krebs-cycle metabolites, phosphorylated glycolytic intermediates, amino acids, peptides, osmolites, nucleosides, organic anions, inorganic anions, iron siderophores, etc. They are found ubiquitously in all three domains of living organisms. Each of the 29 currently recognized families is in general specific for one class of compounds (see http://www-biology.ucsd.edu/~msaier/transport/). While one family in the MFS includes permeases that catalyse amino acid (proline) uptake [the metabolite:H+ symporter (MHS) family (TC 2.A.1.6)], another, the peptide-acetyl-CoA transporter (PAT) family (TC 2.A.1.25), functions in peptide uptake in bacteria (see below) but in acetyl-CoA:CoA antiport in the endoplasmic reticular membrane of higher eukaryotes.

The proton-dependent oligopeptide transporter (POT) family (TC 2.A.17)
The POT family [also called the PTR (peptide transport) family] is primarily a peptide-transporter family, but one member is a nitrate/chlorate permease, and one has been reported to transport histidine as well as nitrate and peptides (Frommer et al., 1994 ; Zhou et al., 1998 ). Because this family is primarily concerned with peptide transport, it will be described below in the section entitled ‘families of peptide transporters’.

The solute:sodium symporter (SSS) family (TC 2.A.21)
All functionally characterized members of the SSS family catalyse solute uptake via Na+ symport (Reizer et al., 1994 ). The solutes transported may be sugars, amino acids, nucleosides, inositols, vitamins, urea or anions, depending on the transport system (Eskandari et al., 1997 ; Prasad et al., 1998 ; Sarker et al., 1997 ). The broad substrate specificity but uniform energy-coupling mechanism exhibited by members of this family are particularly unusual and noteworthy traits of a single family.

Members of the SSS family have been identified in bacteria, archaea and animals. They vary in size from about 400 residues to about 700 residues and possess 12–15 putative TMSs, often sharing a core of 13 TMSs. A 13 TMS topology with a periplasmic N terminus and a cytoplasmic C terminus has been experimentally determined for the proline:Na+ symporter, PutP, of Escherichia coli (Turk & Wright, 1997 ; Jung et al., 1998 ).

The neurotransmitter:sodium symporter (NSS) family (TC 2.A.22)
Members of the large NSS family catalyse uptake of a variety of neurotransmitters, amino acids, osmolytes and related nitrogenous substances by a solute:Na+ symport mechanism (Beckman & Quick, 1998 ; Kavanaugh, 1998 ; Berfield et al., 1999 ). Sometimes Cl- is co-transported, and some of the NSS family members exhibit a K+ dependency. For example, human dopamine, glutamate and {gamma}-aminobutyric acid transporters co-transport their positively charged or zwitterionic substrates with 2 or 3 Na+, and 1 Cl- is probably co-transported, at least with dopamine and {gamma}-aminobutyric acid. The glutamate system may counter-transport K+ (Clark & Amara, 1993 ). Most sequenced members of the NSS family are from animals, and these proteins are generally 600–700 aminoacyl residues in length with 12 TMSs. Bacterial and archaeal homologues have been sequenced, but few functional data are available for these proteins.

Recently, several members of the NSS family have been shown to exhibit channel-like properties under certain experimental conditions. Sizeable unitary ionic currents have been reported for membrane patches containing either the {gamma}-aminobutyrate, noradrenaline or serotonin transporter (Galli et al., 1998 ). Channel-like currents have also been measured for mammalian Na+/H+/K+-coupled glutamate transporters of the dicarboxylate/amino acid:cation symporter (DAACS) family (TC no. 2.A.23; see below). Evidence suggests that these channels can accommodate neurotransmitters as well as inorganic ions. These observations suggest that, as has been demonstrated for carriers of a few other families, neurotransmitter transporters can be manipulated to function as voltage-gated channels. Whether or not this observation is of physiological relevance has yet to be determined.

The DAACS [dicarboxylate/amino acid:cation (Na+ or H+) symporter] family (TC 2.A.23)
Members of the DAACS family catalyse Na+ and/or H+ symport together with (a) a Krebs-cycle dicarboxylate (malate, succinate, or fumarate), (b) a dicarboxylic amino acid (glutamate or aspartate), (c) a small, semipolar, neutral amino acid (Ala, Ser, Cys, Thr) (Arriza et al., 1993 ; Ogawa et al., 1998 ), (d) both neutral and acidic amino acids or (e) most zwitterionic and dibasic amino acids (Reizer et al., 1994 ; Palacín et al., 1998 ; Zarbiv et al., 1998 ). The bacterial members are of about 450 (420–491) aminoacyl residues in length while the mammalian proteins are of about 550 (503–574) residues. These proteins possess between 10 and 12 putative TMSs (Slotboom et al., 1999 ).

All of the bacterial proteins cluster together on the phylogenetic tree as do the mammalian proteins (Saier et al., 1999a ). The mammalian permeases that transport neutral amino acids cluster separately from those that are specific for acidic amino acids. Among the mammalian proteins are neuronal excitatory amino acid neurotransmitter permeases. One of these, the Glt-1 L-glutamate/L-aspartate/D-aspartate transporter co-transports the neurotransmitter with 3 Na+ and 1 H+ and countertransports it against 1 K+ (Clark & Amara, 1993 ).

The MC (mitochondrial carrier) family (TC 2.A.29)
Permease protein subunits of the MC family possess six TMSs and exist in the mitochondrial membrane as homodimers (Aquila et al., 1987 ; Walker & Runswick, 1993 ; Palmieri, 1994 ; Palmieri et al., 1997 , 1999 ; Tzagoloff et al., 1996 ; Echtay et al., 1998 ; Feirmonte et al., 1998 ; Schroers et al., 1998 ). The proteins are of fairly uniform size, about 300 residues (Kuan & Saier, 1993 ; Indiveri et al., 1997 ). They arose by tandem intragenic triplication such that a genetic element encoding two TMSs gave rise to one encoding six TMSs (Saraste & Walker, 1982 ; Walker & Runswick, 1993 ). This event may have occurred when mitochondria first developed their specialized but permanent organellar functions within eukaryotic cells from endosymbiotic bacteria (Kuan & Saier, 1993 ; Indiveri et al., 1997 ). Members of the MC family are found exclusively in eukaryotic organelles although they are nuclearly encoded. Most are found in mitochondria, but some are found in peroxisomes of animals and in amyloplasts of plants (McCammon et al., 1990 ; Sullivan et al., 1991 ; see Kuan & Saier, 1993 for a review). Many of them preferentially catalyse the exchange of one solute for another (antiport); but two of them, the aspartate/glutamate exchanger and the ADP/ATP exchanger can function as anion-selective channels after chemical treatment with thiol reagents (Dierks et al., 1990a , b ). Thirty-four paralogues of the MC family are encoded within the genome of Saccharomyces cerevisiae, and 35 are encoded within the Caenorhabditis elegans genome (Paulsen et al., 1998b ; I. T. Paulsen, S. R. Goldman, W. S. Barnes and M. H. Saier, Jr, unpublished).

The AEC (auxin efflux carrier) family (TC 2.A.69)
Plants possess tissue-specific, PMF-driven, polarly localized cellular auxin efflux systems (Gälweiler et al., 1998 ; Luschnig et al., 1998 ). These carriers are saturable, auxin specific and localized to the basal ends of auxin-transport-competent cells. They may be found in various plant tissues including vascular tissues and roots. They are responsible for the polar (downwards) transport of auxins from the leaves to the roots and function in gravitropism. A single plant such as Arabidopsis thaliana may possess over six genes encoding such systems. Two isoforms, one in vascular tissue (PIN1) and one in roots (REH1) have been functionally characterized as has a homologue from Oryza sativa. These plant proteins are 600–700 aminoacyl residues long and exhibit 8–12 TMSs.

Homologues of the AEC family are found in bacteria (Escherichia coli, Klebsiella pneumoniae, Synechocystis, Aquifex aeolicus, Bacillus subtilis and Rickettsia prowazekii) as well as in archaea (Methanococcus jannaschii and Methanobacterium thermoautotrophicum). The Klebsiella pneumoniae homologue (MdcF, 319 aa) has been suggested to function in malonate uptake (Hoenke et al., 1997 ; Dimroth & Hilbi, 1997 ). The bacterial proteins are generally of 300–400 aa in length.

Yeasts also possess homologues of the AEC family. Saccharomyces cerevisiae has three functionally uncharacterized AEC family members (YL52, spP54072, 64·0 kDa; YNJ5, spP53930, 71·2 kDa; and YB8B, spP38355, 47·5 kDa), and Schizosaccharomyces pombe also has at least one sequenced homologue. It is thus clear that members of the AEC family are widespread, being found in bacteria, archaea, fungi and plants. Caenorhabditis elegans, however, appears to lack identifiable homologues of the AEC family. Based on PSI-BLAST results, the AEC family may be distantly related to the bile acid:Na+ symporter (BASS) family (TC 2.28), which is represented in animals.


   Families of secondary transporters specific for amines, amides and polyamines
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
In addition to the ABC superfamily, which includes members that transport various amines, amides and polyamines, several of the amino acid transporter families described above have been shown to include members that are capable of transporting these compounds. These families include the MFS, APC, BCCT, AAAP, SSS, NSS, MC and AEC families (see Table 2). Additionally, the CAAT family has been implicated in amine transport (Ferguson & Krzycki, 1997 ). Most of these families are described in Table 4, but the BCCT family and the CAAT family, being prokaryote-specific, are described in Table 3. There are no families of secondary carriers in which all members transport simple amines and/or amides exclusively, but not amino acids. Thus, none of the families listed above needs be described here. The fact that most families of permeases which include members that transport amino acids and their derivatives are exclusive for these compounds and do not include members that transport other classes of compounds (i.e. sugars, nucleobases and their derivatives, vitamins, etc.) emphasizes the value of the phylogenetic approach to the functional characterization of recognized permease homologues.


   Families of channel-forming proteins capable of transporting amines and amides
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Three families of channel-forming proteins appear to be capable of transporting simple amines and/or amides. These families are listed in Table 5 and are described below.


View this table:
[in this window]
[in a new window]
 
Table 5. Channel protein families capable of transporting amines and amides

 
The major intrinsic protein (MIP) family (TC 1.A.8)
The MIP family is large and diverse, possessing over 100 sequenced members that all form transmembrane channels. These channel proteins function in the transport of water, small carbohydrates (e.g. glycerol), urea and other small neutral molecules by an energy-independent mechanism (Li et al., 1997 ; Calamita et al., 1998 ; Deen & van Os, 1998 ; Dean et al., 1999 ). They are found ubiquitously in bacteria, archaea and eukaryotes. Phylogenetic clustering of the proteins is largely according to the phylum of the organism of origin, but one to three clusters are observed for each phylogenetic kingdom (plants, animals, yeast, bacteria and archaea) (Park & Saier, 1996 ). The known aquaporins cluster loosely together as do the known glycerol facilitators.

MIP family proteins are believed to form aqueous pores that selectively allow passive transport of their solute(s) across the membrane with minimal apparent recognition (Chrispeels & Maurel, 1994 ; Shukla & Chrispeels, 1998 ). Aquaporins selectively transport water (but not glycerol), while glycerol facilitators selectively transport glycerol but not water (Maurel et al., 1993 ). Glycerol facilitators function as solute nonspecific channels, and may transport glycerol, dihydroxyacetone, propanediol, urea and other small straight-chain neutral molecules in physiologically important processes (Heller et al., 1980 ; Maurel et al., 1994 ). A few members of the family, including the yeast FPS protein (TC 1.A.8.5.1), transport both water and glycerol. Reports of MIP family proteins transporting ions may or may not be physiologically significant. However, demonstration of the involvement of the cyanobacterial channel protein (TC 1.A.8.4.1) in copper homeostasis suggests that it may transport Cu2+ (Kashiwagi et al., 1995 ).

The physiological functions of many MIP family proteins are unknown. They probably consist of homodimers (GlpF of Escherichia coli; TC 1.A.8.1.1) or tetramers (MIP of Bos taurus; TC 1.A.8.8.1). Each subunit spans the membrane six times as putative {alpha}-helices and arose from a three-spanner-encoding genetic element by a tandem, intragenic duplication event (Reizer et al., 1993b ). Consequently, the two halves of the protein are in opposite orientations in the membrane, an unusual feature of a transport protein.

The urea transporter (UT) family (TC 1.A.44)
Members of the UT family are found only in mammals and amphibians (Olives et al., 1994 ; Couriaud et al., 1998 ). In a single species (i.e. rat or human) there may be at least three isoforms. One of the human UT family members is the Kidd (JK) blood group glycoprotein (Lucien et al., 1998 ). Most of these proteins vary in size from 380–400 residues and exhibit 10 putative TMSs, but one of them, a rat urea transporter (gbU77971), is reported to be 929 residues long (Shayakul et al., 1996 ). At least one of these proteins (UT3) can transport water as well as urea (Yang & Verkman, 1998 ). A channel-type mechanism is probable. Two members of this family, UT1 and UT2, may be derived from a single gene by alternative splicing.

The PLM (phospholemman) family (TC 1.A.27)
The PLM family includes mammalian phospholemmans of 8–10 kDa size (Chen et al., 1997 ; Foskett, 1998 ; Kirk & Strange, 1998 ). The human, rat and dog proteins have been sequenced and characterized. They span the membrane once with their N termini outside. These proteins induce a hyperpolarization-activated chloride current in Xenopus oocytes. They are found in muscle and many body tissues and are targets of protein kinases A and C. Other possible members include the chloride-conductance inducer protein, Mat8, and the Na+/K+ ATPase-subunit ‘proteolipid’ (TC 3.A.3). These proteins are smaller, but they exhibit the same orientation in the membrane.

PLM forms anion-selective channels when reconstituted in planar lipid bilayers. These channels display a linear current–voltage relationship, have a unitary conductance and are open most of the time at voltages between -70 and +70 mV. The PLM channel is permeable to both organic and inorganic anions including chloride, taurine, lactate, glutamate, isethionate and gluconate (Kirk & Strange, 1998 ).


   Families of peptide transporters
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
In addition to the ATP-dependent ABC superfamily, in which members can transport peptides with either inward or outward polarity, depending on the family to which the transporter belongs, four families of PMF-driven transporters are primarily concerned with peptide uptake. These families are listed in Table 6 and are described below.


View this table:
[in this window]
[in a new window]
 
Table 6. Secondary active peptide transporter families

 
The PAT (peptide-acetyl-CoA transporter) family of the MFS (TC 2.A.1.25)
Two members of the PAT family of the MFS have been partially characterized from physiological standpoints, but the precise biochemical functions of these proteins are not certain. One of these proteins is the putative acetyl-CoA transporter found in the endoplasmic reticular and golgi membranes of man (Kanamori et al., 1997 ). It is homologous to proteins in Caenorhabditis elegans, Saccharomyces cerevisiae and several Gram-negative bacteria. The other of these proteins, the homologous Escherichia coli AmpG protein, probably brings peptides, including cell-wall degradative peptides, and inducers of ß-lactamase synthesis into the cell (Lindquist et al., 1993 ; Jacobs et al., 1994 ; Park et al., 1998 ). Thus, AmpG may transport cell-wall-derived peptides and glycopeptides. In Haemophilus influenzae, the gene encoding a PAT homologue is found in a gene cluster concerned with lipopolysaccharide synthesis. A homologue from Neisseria gonorrhoeae has also been sequenced. These proteins are of 425–632 aminoacyl residues in length and exhibit 12 putative TMSs as is characteristic of most MFS permeases.

The mechanism of energy coupling exhibited by members of the PAT family is not established, but the topology of these proteins and their established inclusion in the MFS suggest that they are secondary carriers. The acetyl-CoA transporter of mammals is expected to function by acetyl-CoA:CoA antiport while the AmpG protein of Escherichia coli is most likely energized by substrate:H+ symport. Of the PAT family members, prokaryotic proteins are smaller than the eukaryotic proteins by about 100 aminoacyl residues (408–491 residues versus 538–560 residues). Since acetyl-CoA contains several secondary amide (peptide-like) bonds, the inclusion of a substrate such as acetyl-CoA in a family of peptide transporters is not entirely surprising.

Rickettsia prowazekii encodes three AmpG-like paralogues within its small (1·1 Mbp) genome (Andersson et al., 1998 ) although other bacteria (Escherichia coli and Haemophilus influenzae and the two sequenced eukaryotic genomes, Saccharomyces cerevisiae and Caenorhabdidtis elegans), all with much larger genomes, only encode one. Most of the bacteria for which fully sequenced genomes are available, and all of the four archaea with sequenced genomes do not encode a recognizable PAT family member. An analysis of this family has appeared in a recently updated description of the MFS (Saier et al., 1999b ).

The POT (proton-dependent oligopeptide transporter) family (TC 2.A.17)
Proteins of the POT family (Paulsen & Skurray, 1994 ) [also called the PTR (peptide transport) family (Steiner et al., 1995 )] are found in animals, plants, yeast and both Gram-negative and Gram-positive bacteria (Hagting et al., 1994 ; Steiner et al., 1994 ; Daniel, 1996 ; Leibach & Ganapathy, 1996 ; Miyamoto et al., 1996 ; Döring et al., 1998 ; Fei et al., 1998 ). Several of these organisms possess multiple POT family paralogues. The proteins are of about 450–600 aminoacyl residues in length, with the eukaryotic proteins in general being longer than the bacterial proteins (Saier et al., 1999a ). They exhibit 12 putative or established TMSs (Hagting et al., 1997 ; Covitz et al., 1998 ). Some members of the POT family exhibit limited sequence similarity to protein members of the MFS (comparison scores of up to 8 SD for segments in excess of 60 residues in length). Thus the POT family is probably a family within the MFS (Pao et al., 1998 ).

While most members of the POT family catalyse peptide transport, one is a nitrate/chlorate permease (Tsay et al., 1993 ; Frommer et al., 1994 ), and one (or more) can transport histidine as well as nitrate and peptides (Frommer et al., 1994 ; Zhou et al., 1998 ). Some of the peptide transporters can also transport antibiotics. They function by proton symport, but the substrate:H+ stoichiometry is variable: the high-affinity rat PepT2 carrier catalyses uptake of 2 and 3 H+ with neutral and anionic dipeptides, respectively, while the low-affinity PepT1 carrier catalyses uptake of 1 H+ per neutral peptide (Chen et al., 1999 ).

The OPT (oligopeptide transporter) family (TC 2.A.67)
The OPT family consists of transporters for oligopeptides of 4–6 aminoacyl residues (Lubkowitz et al., 1997 , 1998 ). Two transporters from Saccharomyces cerevisiae, one from Schizosaccharomyces pombe and one from Candida albicans have been functionally characterized, and all are peptide-uptake systems. Saccharomyces cerevisiae possesses three paralogues of the OPT family while Schizosaccharomyces pombe has at least two. One of the Schizosaccharomyces pombe homologues is the sexual differentiation process (ISP4) protein. Homologues are also found in plants, and distant homologues may be present in bacteria and archaea as well. The prokaryotic homologues are very distant, being revealed only upon PSI-BLAST iterations, and they are uncharacterized functionally. Energy coupling probably involves H+ symport. The full-length yeast proteins are reported to be 700–900 residues long and exhibit up to 12 TMSs. A putative bacterial homologue from Haemophilus influenzae is 633 aminoacyl residues long and exhibits 15 putative TMSs.

The PUP (peptide-uptake permease) family (TC 9.A.18)
Two partially functionally characterized proteins, the SbmA protein (406 aa) of Escherichia coli and the BacA protein (420 aa) of Rhizobium meliloti, define the PUP family (Glazebrook et al., 1993 ; Salomón & Farías, 1995 ; Ichige & Walker, 1997 ). SbmA catalyses uptake of thiazole ring-containing peptide antibiotics such as microcin B17 and microcin J25 as well as the non-peptide antibiotic, bleomycin. BacA is a nodulation protein essential for bacterial development when Rhizobium is in symbiosis with a leguminous plant such as alfalfa. These two proteins exhibit 64% identity and are functionally interchangeable in both Escherichia coli and Rhizobium meliloti. Rhizobium meliloti bacA null mutants show increased resistance to bleomycin and certain aminoglycosides as well as increased sensitivity to ethanol and detergents. The latter properties are not characteristic of Escherichia coli sbmA mutants. It is hypothesized that BacA may take up peptide substances required for developmental progression towards bacteroid formation. Based on the mutant analyses, BacA (but not SbmA) may also play a role in the maintenance of membrane integrity.

Proteins of the PUP family are homologous, but distantly related, to a few putative ABC-type transporters of Gram-negative and Gram-positive bacteria. Unlike SbmA and BacA, the latter proteins possess ABC-containing domains. SbmA and BacA also differ from these putative ABC proteins in possessing seven rather than six putative TMSs per polypeptide chain. It is possible that ATP-hydrolysing protein constituents (ABC proteins) of the PUP family transporters will be found. If so, these systems may prove to be energized by ATP hydrolysis. However, the mechanism of energy coupling to PUP family permeases is currently unknown. It is for this reason that the PUP family is placed into the ‘9’ category of the TC system. If ABC proteins prove not to be associated with these permeases, they will provide one of the few examples of secondary carriers that are phylogenetically related to primary active-transport systems.


   Protein-transport systems
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Twenty transporter families are known to include members that function in the transmembrane transport of proteins (Table 7). Many of these (TC category 3.A) are primary active-transport systems driven by either ATP or GTP hydrolysis although protein export via several of these systems is stimulated by the PMF. One family (twin arginine targeting translocase, Tat; TC 2.A.64) exports proteins in a process that is energized exclusively by the PMF, implying that protein export is coupled to proton import. Transport systems of the Tat family therefore fall into the category of secondary transporters. Finally, many channel-forming proteins (categories IX and X in Table 7) have the capacity to transport proteins, most of them probably in an energy-independent fashion. These families will not be described here but the interested reader will find descriptions of these families at http://www-biology.ucsd.edu/~msaier/transport/.


View this table:
[in this window]
[in a new window]
 
Table 7. Protein secretory pathways (PSP) in living organisms

 

   Conclusion
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Over two dozen families of transporters are currently known to be responsible for the transmembrane transport of amino acids, small amines, polyamines, amides and peptides. Three of these are families of channel-forming proteins, and they generally transport urea and other small nitrogen-containing compounds. Of the 21 currently recognized families of secondary carriers specific for amino acids and their derivatives, 11 are found only in prokaryotes. Nine of these families include members that have so far been found only in bacteria, but two are also represented in archaea. All but one of the prokaryote-specific transporter families are capable of transporting amino acids, but the one exceptional family consists of transporters that seem to be highly selective for peptides and glycopeptides. An additional ten families are either ubiquitous (eight families) or restricted to eukaryotes (two families). Eight of the 19 amino acid-transporting families of secondary carriers include members that also transport small amines and amides, and two of these families include members that exhibit the capacity to transport peptides. Only two families include members that appear to transport peptides specifically, lacking the capacity to transport simple amino acids.

Numerous permeases within the ABC superfamily catalyse either uptake or extrusion of amino acids, amines and/or peptides (Table 1). For the uptake of amino acids, one family (TC 3.A.1.3) is generally selective for polar amino acids while a second (TC 3.A.1.4) is selective for nonpoplar amino acids. Only one uptake family (TC 3.A.1.5) within the ABC superfamily includes members that transport peptides. Three families include members that take up polyamines (TC 3.A.1.11), quaternary amines (TC 3.A.1.12) and taurine (TC 3.A.1.17). Amines are often the decarboxylation products of amino acids or the methylated derivatives of these products, while peptides and depsipeptides (peptide-like molecules synthesized by enzymes in nonribosome-dependent processes) are condensation products of amino acids. All of these compounds can therefore be thought of as amino acid derivatives.

The ABC uptake permeases segregate on a phylogenetic tree from the efflux systems (Saurin et al., 1999 ). Of the efflux systems (Table 1), the peptide transporters are found primarily in seven families, five prokaryote-specific families (TC 3.A.1.111–113, 116, 118) and two eukaryote-specific families (TC 3.A.1.206 and 3.A.1.208). Two ABC families (TC 3.A.1.109 and 110) function in the secretion of proteins from bacteria, but no ABC-type eukaryotic exporters have been shown to function in protein secretion. This may be due to the fact that most proteins and complex carbohydrates are secreted in eukaryotes primarily, if not exclusively, by exocytosis. We note, therefore that of the 48 recognized families of ABC transporters, 15, or nearly one-third of all recognized ABC transport families, are primarily concerned with transport of amino acids and their derivatives. Taken together, about 10% of all currently recognized families of transporters include members that are known to transport low-molecular-mass amines, amides, amino acids and peptides, with another 10% functioning in the export of proteins. These large percentages illustrate the importance of amino acids and their derivatives to the normal function of all biological cells.


   ACKNOWLEDGEMENTS
 
I wish to particularly thank my colleague, Dr Ian Paulsen, who directed all of the genome analyses presented on our web site (http://www-biology.ucsd.edu/~msaier/transport/) whose work provided the initial basis for formulation of the TC system, and whose numerous discussions have been of tremendous intellectual benefit. I also acknowledge the many students whose phylogenetic analyses were described in this review. Dr Arnost Kotyk, Chairman of the Transport Protein Nomenclature Panel of the International Union of Biochemistry and Molecular Biology (IUBMB), as well as the other members of this panel (Rolf Apweiler, Amos Bairoch, Andre Lupas, Ian Paulsen and myself) are gratefully acknowledged for their valuable suggestions. Useful suggestions were also provided by André Goffeau and Peter Karp, members of the IUBMB nomenclature panel in absentia. The work described in this review was supported by NIH grants 2R01 AI14176 from The National Institute of Allergy and Infectious Diseases and 9RO1 GM55434 from the National Institute of General Medical Sciences, as well as the M. H. Saier, Sr Memorial Research Fund.


   REFERENCES
TOP
Overview
Background
Characteristics of transporters...
Most amino acid transporters...
ABC (ATP-binding cassette) -type...
Secondary carriers
Secondary amino acid transporter...
Amino acid transporter families...
Families of secondary...
Families of channel-forming...
Families of peptide transporters
Protein-transport systems
Conclusion
REFERENCES
 
Aleshin, V. V., Zakataeva, N. P. & Livshits, V. A. (1999). A new family of amino acid efflux proteins.Trends Biochem Sci24, 133-135.[Medline]

Andersson, S. G. E., Zomorodipour, A., Andersson, J. O. & 7 other authors (1998). The genome sequence of Rickettsia prowazekii and the origin of mitochondria. Nature 396, 133–140.[Medline]

Aquila, H., Link, T. A. & Klingenberg, T. (1987). Solute carriers involved in energy transfer of mitochondria form a homologous protein family.FEBS Lett212, 1-9.[Medline]

Arriza, J. L., Kavanaugh, M. P., Fairman, W. A., Wu, Y. N., Murdoch, G. H., North, R. A. & Amara, S. G. (1993). Cloning and expression of a human neutral amino acid transporter with structural similarity to the glutamate transporter gene family.J Biol Chem268, 15329-15332.[Abstract/Free Full Text]

Beckman, M. L. & Quick, M. W. (1998). Neurotransmitter transporters: regulators of function and functional regulation.J Membr Biol164, 1-10.[Medline]

Bennett, M. J., Marchant, A., Green, H. G., May, S. T., Ward, S. P., Millner, P. A., Walker, A. R., Schulz, B. & Feldmann, K. A. (1996). Arabidopsis AUX1 gene: a permease-like regulator of root gravitropism. Science273, 948-950.[Abstract]

Berfield, J. L., Wang, L. C. & Reith, M. E. A. (1999). Which form of dopamine is the substrate for the human dopamine transporter: the cationic or the uncharged species?J Biol Chem274, 4876-4882.[Abstract/Free Full Text]

Berg, M., Hilbi, H. & Dimroth, P. (1997). Sequence of a gene cluster from Malonomonas rubra encoding components of the malonate decarboxylase Na+ pump and evidence for their function.Eur J Biochem245, 103-105.[Abstract]

Brechtel, C. E. & King, S. C. (1998). 4-aminobutyrate (GABA) transporters from the amine-polyamine-choline superfamily: substrate specificity and ligand recognition profile of the 4-aminobutyrate permease from Bacillus subtilis.Biochem J333, 565-571.[Medline]

Bröer, S. & Krämer, R. (1991a). Lysine excretion by Corynebacterium glutamicum. I. Identification of a specific secretion carrier system.Eur J Biochem202, 131-135.[Abstract]

Bröer, S. & Krämer, R. (1991b). Lysine excretion by Corynebacterium glutamicum. II. Energetics and mechanism of the transport system.Eur J Biochem202, 137-143.[Abstract]

Calamita, G., Kempf, B., Bonhivers, M., Bishai, W. R., Bremer, E. & Agre, P. (1998). Regulation of the Escherichia coli water channel gene aqpZ.Proc Natl Acad Sci USA95, 3627-3631.[Abstract/Free Full Text]

Chen, L. S. K., Lo, C. F., Numann, R. & Cuddy, M. (1997). Characterization of the human and rat phospholemman (PLM) cDNAs and localization of the human PLM gene to chromosome 19q13.1.Genomics41, 435-443.[Medline]

Chen, X.-Z., Zhu, T., Smith, D. E. & Hediger, M. A. (1999). Stoichiometry and kinetics of the high-affinity H+-coupled peptide transporter PepT2.J Biol Chem274, 2773-2779.[Abstract/Free Full Text]

Chrispeels, M. J. & Maurel, C. (1994). Aquaporins: the molecular basis of facilitated water movement through living plant cells?Plant Physiol105, 9-13.[Free Full Text]

Clark, J. A. & Amara, S. G. (1993). Amino acid neurotransmitter transporters: structure, function, and molecular diversity.Bioessays15, 323-332.[Medline]

Closs, E. I., Albritton, L. M., Kim, J. W. & Cunningham, J. M. (1993). Identification of a low affinity, high capacity transporter of cationic amino acids in mouse liver.J Biol Chem268, 7538-7544.[Abstract/Free Full Text]

Cosgriff, A. J. & Pittard, A. J. (1997). A topological model for the general aromatic amino acid permease, AroP, of Escherichia coli.J Bacteriol179, 3317-3323.[Abstract]

Couriaud, C., Ripoche, P. & Rousselet, G. (1998). Cloning and functional characterization of a rat urea transporter expression in the brain.Biochim Biophys Acta1309, 197-199.

Covitz, K.-M. Y., Amidon, G.-L. & Sadée, W. (1998). Membrane topology of the human dipeptide transporter, hPEPT1, determined by epitope insertions.Biochemistry37, 15214-15221.[Medline]

Daniel, H. (1996). Function and molecular structure of brush border membrane peptide/H+ symporters. J Membr Biol154, 197-203.[Medline]

Dean, R. M., Rivers, R. L., Zeidel, M. L. & Roberts, D. M. (1999). Purification and functional reconstitution of soybean nodulin 26: an aquaporin with water and glycerol transport properties.Biochemistry38, 347-353.[Medline]

Deen, P. M. T. & van Os, C. H. (1998). Epithelial aquaporins.Curr Opin Cell Biol10, 435-442.[Medline]

Deguchi, Y., Yamato, I. & Anraku, Y. (1990). Nucleotide sequence of gltS, the Na+/glutamate symport carrier gene of Escherichia coli B.J Biol Chem265, 21704-21708.[Abstract/Free Full Text]

Didion, T., Regenberg, B., Jørgensen, M. U., Kielland-Brandt, M. C. & Andersen, H. A. (1998). The permease homologue Ssy1p controls the expression of amino acid and peptide transporter genes in Saccharomyces cerevisiae. Mol Microbiol27, 643-650.[Medline]

Dierks, T., Salentin, A., Heberger, C. & Krämer, R. (1990a). The mitochondrial aspartate/glutamate and ADP/ATP carriers switch from obligate counterexchange to unidirectional transport after modification by SH-reagents.Biochim Biophys Acta1028, 268-280.[Medline]

Dierks, T., Salentin, A. & Krämer, R. (1990b). Pore-like and carrier-like properties of the mitochondrial aspartate/glutamate carrier after modification by SH-reagents: evidence for a preformed channel as a structural requirement of carrier-mediated transport.Biochim Biophys Acta1028, 281-288.[Medline]

Dimroth, P. & Hilbi, H. (1997). Enzymatic and genetic basis for bacterial growth on malonate.Mol Microbiol25, 3-10.[Medline]

Döring, F., Will, J., Amasheh, S., Clauss, W., Ahlbrecht, H. & Daniel, H. (1998). Minimal molecular determinants of substrates for recognition by the intestinal peptide transporter.J Biol Chem273, 23211-23218.[Abstract/Free Full Text]

Echtay, K. S., Bienengraeber, M., Winkler, E. & Klingenberg, M. (1998). In the uncoupling protein (UCP-1) His-214 is involved in the regulation of purine nucleoside triphosphate but not diphosphate binding.J Biol Chem273, 24368-24374.[Abstract/Free Full Text]

Eichler, K., Bourgis, F., Buchet, A., Kleber, H. P. & Mandrand-Berthelot, M. A. (1994). Molecular characterization of the cai operon necessary for carnitine metabolism in Escherichia coli.Mol Microbiol13, 775-786.[Medline]

Engel, P., Krämer, R. & Unden, G. (1994). Transport of C4-dicarboxylates by anaerobically grown Escherichia coli: energetics and mechanism of exchange, uptake and efflux. Eur J Biochem222, 605-614.[Abstract]

Eskandari, S., Loo, D. D. F., Dai, G., Levy, O., Wright, E. M. & Carrasco, N. (1997). Thyroid Na+/I- symporter: mechanism, stoichiometry, and specificity.J Biol Chem272, 27230-27238.[Abstract/Free Full Text]

Farcasanu, I. C., Mizunuma, M., Hirata, D. & Miyakawa, T. (1998). Involvement of histidine permease (Hip1p) in manganese transport in Saccharomyces cerevisiae. Mol Gen Genet259, 541-548.[Medline]

Fei, Y.-J., Fujita, T., Lapp, D. F., Ganapathy, V. & Leibach, F. H. (1998). Two oligopeptide transporters from Caenorhabditis elegans: molecular cloning and functional expression.Biochem J322, 565-572.

Feirmonte, G., Palmieri, L., Dolce, V., Lasorsa, F. M., Palmieri, F., Runswick, M. J. & Walker, J. E. (1998). The sequence, bacterial expression, and functional reconstitution of the rat mitochondrial dicarboxylate transporter cloned via distant homologs in yeast and Caenorhabditis elegans.J Biol Chem273, 24754-24759.[Abstract/Free Full Text]

Fekkes, P. & Driessen, A. J. M. (1999). Protein targeting to the bacterial cytoplasmic membrane. Microbiol Mol Biol Rev63, 161-173.[Abstract/Free Full Text]

Ferguson, D. J. & Krzycki, J. A. (1997). Reconstruction of trimethylamine-dependent coenzyme M methylation with the trimethylamine corrinoid protein and the isozymes of methyltransferase II from Methanosarcina barkeri.J Bacteriol179, 846-852.[Abstract]

Fischer, W.-N., Kwart, M., Hummel, S. & Frommer, W. B. (1995). Substrate specificity and expression profile of amino acid transporters (AAPs) in Arabidopsis.J Biol Chem270, 16315-16320.[Abstract/Free Full Text]

Forward, J., Behrendt, M. C., Wyborn, N. R., Cross, R. & Kelly, D. J. (1997). TRAP transporters: a new family of periplasmic solute transport systems encoded by the dctPQM genes of Rhodobacter capsulatus and by homologs in diverse Gram-negative bacteria.J Bacteriol179, 5482-5493.[Abstract]

Foskett, J. K. (1998). ClC and CFTR chloride channel gating.Annu Rev Physiol60, 689-717.[Medline]

Frommer, W. B., Hummel, S. & Rentsch, D. (1994). Cloning of an Arabidopsis histidine transporting protein related to nitrate and peptide transporters.FEBS Lett347, 185-189.[Medline]

Fuqua, C., Winans, S. C. & Greenberg, E. P. (1996). Census and consensus in bacterial ecosystems: the LuxR–LuxI family of quorum-sensing transcriptional regulators.Annu Rev Microbiol50, 727-751.[Medline]

Galli, A., Blakely, R. D. & DeFelice, L. J. (1998). Patch-clamp and amperometric recordings from norepinephrine transporters: channels activity and voltage-dependent uptake. Proc Natl Acad Sci USA 13260–13265.

Gälweiler, L., Guan, C., Müller, A., Wisman, E., Mendgen, K., Yephremov, A. & Palme, K. (1998). Regulation of polar auxin transport by AtPIN1 in Arabidopsis vascular tissue.Science282, 2226-2230.[Abstract/Free Full Text]

Glazebrook, J., Ichige, A. & Walker, G. C. (1993). A Rhizobium meliloti homolog of the Escherichia coli peptide-antibiotic transport protein SbmA is essential for bacteroid development.Genes Dev7, 1485-1497.[Abstract]

Golby, P., Kelly, D. J., Guest, J. R. & Andrews, S. C. (1998). Topological analysis of DcuA, an anaerobic C4-dicarboxylate transporter of Escherichia coli.J Bacteriol180, 4821-4827.[Abstract/Free Full Text]

Goss, T. J., Schweizer, H. P. & Datta, P. (1988). Molecular characterization of the tdc operon of Escherichia coli K-12.J Bacteriol170, 5352-5359.[Medline]

Hagting, A., Kunji, E. R. S., Leenhouts, K. J., Poolman, B. & Konings, W. N. (1994). The di- and tripeptide transport protein of Lactococcus lactis. J Biol Chem269, 11391-11399.[Abstract/Free Full Text]

Hagting, A., van der Velde, J., Poolman, B. & Konings, W. N. (1997). Membrane topology of the di- and tripeptide transport protein of Lactococcus lactis.Biochemistry36, 6777-6785.[Medline]

von Heijne, G. (1992). Membrane protein structure prediction: hydrophobicity analysis and positive-inside rule. J Mol Biol225, 487-494.[Medline]

Heller, K. B., Lin, E. C. C. & Wilson, T. H. (1980). Substrate specificity and transport properties of the glycerol facilitator of Escherichia coli.J Bacteriol144, 274-278.[Medline]

Hoenke, S., Schmid, M. & Dimroth, P. (1997). Sequence of a gene cluster from Klebsiella pneumoniae encoding malonate decarboxylase and expression of the enzyme in Escherichia coli.Eur J Biochem246, 530-538.[Abstract]

Hu, L. A. & King, S. C. (1998a). Functional significance of the ‘signature cysteine’ in helix 8 of the Escherichia coli 4-aminobutyrate transporter from the amine-polyamine-choline superfamily.J Biol Chem273, 20162-20167.[Abstract/Free Full Text]

Hu, L. A. & King, S. C. (1998b). Functional sensitivity of polar surfaces on transmembrane helix 8 and cytoplasmic loop 8–9 of the Escherichia coli GABA (4-aminobutyrate) transporter encoded by gabP: mutagenic analysis of a consensus amphipathic region found in transporters from bacteria to mammals.Biochem J330, 771-776.[Medline]

Hu, L. A. & King, S. C. (1998c). Membrane topology of the Escherichia coli {gamma}-aminobutyrate transporter: implications on the topology and mechanism of prokaryotic and eukaryotic transporters from the APC superfamily.Biochem J336, 69-76.[Medline]

Ichige, A. & Walker, G. C. (1997). Genetic analysis of the Rhizobium meliloti bacA gene: functional interchangeability with the Escherichia coli sbmA gene and phenotypes of mutants.J Bacteriol179, 209-216.[Abstract]

Indiveri, C., Iacobazzi, V., Giangregorio, N. & Palmieri, F. (1997). The mitochondria carnitine carrier protein: cDNA cloning, primary structure and comparison with other mitochondrial transport proteins.Biochem J321, 713-719.[Medline]

Isnard, A. D., Thomas, D. & Surdin-Kerjan, Y. (1996). The study of methionine uptake in Saccharomyces cerevisiae reveals a new family of amino acid permeases.J Mol Biol262, 473-484.[Medline]

Jack, D. L., Paulsen, I. T. & Saier, M. H.Jr (2000). The APC superfamily of transporters specific for amino acids, polyamines and organocations.Microbiology146, 1797-1814.[Abstract/Free Full Text]

Jacobs, C., Huang, L., Bartowsky, E., Normark, S. & Park, J. T. (1994). Bacterial cell wall recycling provides cystolic muropeptides as effectors for ß-lactamase induction.EMBO J13, 4684-4694.[Abstract]

Jacobs, M. H. J., van der Heide, T., Driessen, A. J. M. & Konings, W. N. (1996). Glutamate transport in Rhodobacter sphaeroides is mediated by a novel binding protein-dependent secondary transport system.Proc Natl Acad Sci USA93, 12786-12790.[Abstract/Free Full Text]

Jung, H., Rübenhagen, R., Tebbe, S., Leifker, K., Tholema, N., Quick, M. & Schmid, R. (1998). Topology of the Na+/proline transporter of Escherichia coli.J Biol Chem273, 26400-26407.[Abstract/Free Full Text]

Kanamori, A., Nakayama, J., Fukuda, M. N., Stallcup, W. B., Sasaki, K., Fukuda, M. & Hirabayashi, Y. (1997). Expression, cloning, and characterization of a cDNA encoding a novel membrane protein required for the formation of O-acetylated ganglioside: a putative acetyl-CoA transporter.Proc Natl Acad Sci USA94, 2897-2902.[Abstract/Free Full Text]

Kappes, R., Kempf, B. & Bremer, E. (1996). Three transport systems for the osmoprotectant glycine betaine operate in Bacillus subtilis: characterization of OpuD.J Bacteriol178, 5071-5079.[Abstract]

Kashiwagi, S., Kanamaru, K. & Mizuno, T. (1995). A Synechococcus gene encoding a putative pore-forming intrinsic membrane protein. Biochim Biophys Acta1237, 189-192.[Medline]

Kashiwagi, K., Shibuya, S., Tomitori, H., Kuraishi, A. & Igaragshi, K. (1997). Excretion and uptake of putrescine by the PotE protein in Escherichia coli. J Biol Chem272, 6318-6323.[Abstract/Free Full Text]

Kavanaugh, M. P. (1998). Neurotransmitter transport: models in flux.Proc Natl Acad Sci USA95, 12737-12738.[Free Full Text]

Kempf, B. & Bremer, E. (1998). Uptake and synthesis of compatible solutes as microbial stress responses to high-osmolality environments. Arch Microbiol170, 319-330.[Medline]

Kirk, K. & Strange, K. (1998). Functional properties and physiological roles of organic solute channels.Annu Rev Physiol60, 719-739.[Medline]

Knutson, V. P. (1991). Cellular trafficking and processing of the insulin receptor.FASEB J5, 2130-2138.[Abstract/Free Full Text]

Koch, H.-G., Hengelage, T., Neumann-Haefelin, C., MacFarlane, J., Hoffschulte, H. K., Schimz, K.-L., Mechler, B. & Müller, M. (1999). In vitro studies with purified components reveal signal recognition particle (SRP) and SecA/SecB as constituents of two independent protein-targeting pathways of Escherichia coli.Mol Biol Cell10, 2163-2173.[Abstract/Free Full Text]

Kuan, J. & Saier, M. H.Jr (1993). The mitochondrial carrier family of transport proteins: structural, functional and evolutionary relationships.Crit Rev Biochem Mol Biol28, 209-233.[Abstract]

Lamark, T., Kaasen, I., Eshoo, M. W., Falkenberg, P., McDougall, J. & Strom, A. R. (1991). DNA sequence and analysis of the bet genes encoding the osmoregulatory choline-glycine betaine pathway of Escherichia coli.Mol Microbiol5, 1049-1064.[Medline]

Leibach, F. H. & Ganapathy, V. (1996). Peptide transporters in the intestine and the kidney.Annu Rev Nutr16, 99-119.[Medline]

Li, H., Lee, S. & Jap, B. K. (1997). Molecular design of aquaporin-1 water channel as revealed by electron crystallography.Nat Struct Biol4, 263-265.[Medline]

Lindquist, S., Weston-Hafer, K., Schmidt, H., Pul, C., Korfmann, G., Erickson, J., Sanders, C., Martin, H. H. & Normark, S. (1993). AmpG, a single transducer in chromosomal ß-lactamase induction.Mol Microbiol9, 703-715.[Medline]

Lubkowitz, M. A., Hauser, L., Breslav, M., Naider, F. & Becker, J. M. (1997). An oligopeptide transport gene from Candida albicans.Microbiology143, 387-396.[Abstract]

Lubkowitz, M. A., Barnes, D., Breslav, M., Burchfield, A., Naider, F. & Becker, J. M. (1998). Schizosaccharomyces pombe isp4 encodes a transporter representing a novel family of oligopeptide transporters.Mol Microbiol28, 729-741.[Medline]

Lucien, N., Sidoux-Walter, F., Olives, B., Moulds, J., Le Pennec, P.-Y., Cartron, J.-P. & Bailly, P. (1998). Characterization of the gene encoding the human Kidd blood group/urea transporter protein. J Biol Chem273, 12973-12980.[Abstract/Free Full Text]

Luschnig, C., Gaxiola, R. A., Grisafi, P. & Fink, G. R. (1998). EIR1, a root-specific protein involved in auxin transport, is required for gravitropism in Arabidopsis thaliana.Genes Dev12, 2175-2187.[Abstract/Free Full Text]

McCammon, M. T., Dowds, C. A., Orth, K., Moomaw, C. R., Slaughter, C. A. & Goodman, J. M. (1990). Sorting of peroxisomal membrane protein PMP47 from Candida boidinii into peroxisomal membranes of Saccharomyces cerevisiae. J Biol Chem265, 20098-20105.[Abstract/Free Full Text]

McIntire, S. L., Reimer, R. J., Schuske, K., Edwards, R. H. & Jorgensen, E. M. (1997). Identification and characterization of the vesicular GABA transporter.Nature389, 870-876.[Medline]

Mastroberardino, L., Spindler, B., Pfeiffer, R., Skelly, P. J., Loffing, J., Shoemaker, C. B. & Verrey, F. (1998). Amino-acid transport by heterodimers of 4F2hc/CD98 and members of a permease family.Nature395, 288-291.[Medline]

Maurel, C., Reizer, J., Schroeder, J. I. & Chrispeels, M. J. (1993). The vacuolar membrane protein gamma-TIP creates water-specific channels in Xenopus oocytes.EMBO J12, 2241-2247.[Abstract]

Maurel, C., Reizer, J., Schroeder, J. I., Chrispeels, M. J. & Saier, M. H.Jr (1994). Functional characterization of the Escherichia coli glycerol facilitator, GlpF, in Xenopus oocytes.J Biol Chem269, 11869-11872.[Abstract/Free Full Text]

Miyamoto, K.-I., Shiraga, T., Morita, K. & 7 other authors (1996). Sequence, tissue distribution and developmental changes in rat intestinal oligopeptide transporter. Biochim Biophys Acta 1305, 34–38.[Medline]

Ogawa, W., Kim, Y. M., Mizushima, T. & Tsuchiya, T. (1998). Cloning and expression of the gene for the Na+-coupled serine transporter from Escherichia coli, and characteristics of the transporter.J Bacteriol180, 6749-6752.[Abstract/Free Full Text]

Olives, B., Neau, P., Bailly, P., Hediger, M. A., Rousselet, G., Cartron, J. P. & Ripoche, P. (1994). Cloning and functional expression of a urea transporter from human bone marrow cells.J Biol Chem269, 31649-31652.[Abstract/Free Full Text]

Palacín, M., Estévez, R., Bertran, J. & Zorzano, A. (1998). Molecular biology of mammalian plasma membrane amino acid transporters.Physiol Rev78, 969-1054.[Abstract/Free Full Text]

Palmieri, F. (1994). Mitochondrial carrier proteins. FEBS Lett346, 48-54.[Medline]

Palmieri, L., Lasorsa, F. M., De Palma, A., Palmieri, F., Runswick, M. J. & Walker, J. E. (1997). Identification of the yeast ACR1 gene product as a succinate-fumarate transporter essential for growth on ethanol or acetate.FEBS Lett417, 114-118.[Medline]

Palmieri, L., Vozza, A., Hönlinger, A., Dietmeier, K., Palmisano, A., Zara, V. & Palmieri, F. (1999). The mitochondrial dicarboxylate carrier is essential for the growth of Saccharomyces cerevisiae on ethanol or acetate as the sole carbon source.Mol Microbiol31, 569-577.[Medline]

Pao, S. S., Paulsen, I. T. & Saier, M. H.Jr (1998). The major facilitator superfamily.Microbiol Mol Biol Rev62, 1-32.[Abstract/Free Full Text]

Park, J. H. & Saier, M. H.Jr (1996). Phylogenetic characterization of the MIP family of transmembrane channel proteins. J Membr Biol153, 171-180.[Medline]

Park, J. T., Raychaudhuri, D., Li, H., Normark, S. & Mengin-Lecreulx, D. (1998). MppA, a periplasmic binding protein essential for import of the bacterial cell wall peptide L-alanyl-{gamma}-D-glutamyl-meso-diaminopimelate.J Bacteriol180, 1215-1223.[Abstract/Free Full Text]

Paulsen, I. T. & Skurray, R. A. (1994). The POT family of transport proteins.Trends Biochem Sci18, 404.

Paulsen, I. T., Sliwinski, M. K. & Saier, M. H.Jr (1998a). Microbial genome analyses: global comparisons of transport capabilities based on phylogenies, bioenergetics and substrate specificities. J Mol Biol277, 573-592.[Medline]

Paulsen, I. T., Sliwinski, M. K., Nelissen, B., Goffeau, A. & Saier, M. H.Jr (1998b). Unified inventory of established and putative transporters encoded within the complete genome of Saccharomyces cerevisiae.FEBS Lett430, 116-125.[Medline]

Peter, H., Weil, B., Burkovski, A., Krämer, R. & Morbach, S. (1998). Corynebacterium glutamicum is equipped with four secondary carriers for compatible solutes: identification, sequencing, and characterization of the proline/ectoine uptake system, ProP, and the ectoine/proline/glycine betaine carrier, EctP.J Bacteriol180, 6005-6012.[Abstract/Free Full Text]

Prasad, P. D., Wang, H., Kekuda, R., Fujita, T., Fei, Y.-J., Devoe, L. D., Leibach, F. H. & Ganapathy, V. (1998). Cloning and functional expression of a cDNA encoding a mammalian sodium-dependent vitamin transporter mediating the uptake of pantothenate, biotin, and lipoate. J Biol Chem273, 7501-7506.[Abstract/Free Full Text]

Rabus, R., Jack, D. L., Kelly, D. J. & Saier, M. H.Jr (1999). TRAP transporters: an ancient family of extracytoplasmic solute receptor-dependent secondary active transporters.Microbiology145, 3431-3445.[Abstract/Free Full Text]

Reizer, J., Finley, K., Kakuda, D., MacLeod, C. L., Reizer, A. & Saier, M. H.Jr (1993a). Mammalian integral membrane receptors are homologous to facilitators and antiporters of yeast, fungi, and eubacteria.Protein Sci2, 20-30.[Abstract/Free Full Text]

Reizer, J., Reizer, A. & Saier, M. H.Jr (1993b). The MIP family of integral membrane channel proteins: sequence comparisons, evolutionary relationships, reconstructed pathway of evolution and proposed functional differentiation of the two repeated halves of the proteins.Crit Rev Biochem Mol Biol28, 235-257.[Abstract]

Reizer, J., Michotey, V., Reizer, A. & Saier, M. H.Jr (1994). Novel phosphotransferase system genes revealed by bacterial genome analysis: unique, putative fructose- and glucoside-specific systems. Protein Sci3, 440-450.[Abstract/Free Full Text]

Reizer, J., Charbit, A., Reizer, A. & Saier, M. H.Jr (1996). Novel phosphotransferase system genes revealed by bacterial genome analysis: operons encoding homologues of sugar-specific permease domains of the phosphotransferase system and pentose catabolic enzymes. Genome Sci Tech1, 53-75.

Rentsch, D., Hirner, B., Schmeizer, E. & Frommer, W. B. (1996). Salt stress-induced proline transporters and salt stress-repressed broad specificity amino acid permeases identified by suppression of a yeast amino acid permease-targeting mutant.Plant Cell8, 1437-1446.[Abstract/Free Full Text]

Reverchon, S., Nasser, W. & Robert-Baudouy, J. (1994). pecS: a locus controlling pectinase, cellulase and blue pigment production in Erwinia chrysanthemi.Mol Microbiol11, 1127-1139.[Medline]

Saier, M. H.Jr (1994). Computer-aided analyses of transport protein sequences: gleaning evidence concerning function, structure, biogenesis, and evolution.Microbiol Rev58, 71-93.[Abstract]

Saier, M. H.Jr (1996). Phylogenetic approaches to the identification and characterization of protein families and superfamilies.Microb Comp Genomics1, 129-150.[Medline]

Saier, M. H.Jr (1998). Molecular phylogeny as a basis for the classification of transport proteins from bacteria, archaea and eukarya. In Advances in Microbial Physiology, pp. 81-136. Edited by R. K. Poole. San Diego, CA: Academic Press.

Saier, M. H.Jr (1999a). Classification of transmembrane transport systems in living organisms. In Biomembrane Transport, pp. 265-276. Edited by L. Van Winkle. San Diego, CA: Academic Press.

Saier, M. H.Jr (1999b). Eukaryotic transmembrane solute transport systems. In International Review of Cytology: a Survey of Cell Biology, pp. 61-136. Edited by K. W. Jeon. San Diego, CA: Academic Press.

Saier, M. H.Jr (1999c). Genome archeology leading to the characterization and classification of transport proteins.Curr Opin Microbiol2, 555-561.[Medline]

Saier, M. H.Jr & Tseng, T.-T. (1999). Evolutionary origins of transmembrane transport systems. In Transport of Molecules Across Microbial Membranes (Symposium no. 58, Society for General Microbiology), pp. 252-274. Edited by J. K. Broome-Smith, S. Baumberg, C. J. Stirling & F. B. Ward. Cambridge: Cambridge University Press.

Saier, M. H.Jr, Müller, M. & Werner, P. K. (1989). Insertion of proteins into bacterial membranes: mechanism, characteristics, and comparisons with the eucaryotic process.Microbiol Rev53, 333-336.

Saier, M. H., Jr, Eng, B. H., Fard, S. & 15 other authors (1999a). Phylogenetic characterization of novel transport protein families revealed by genome analyses. Biochim Biophys Acta 1422, 1–56.[Medline]

Saier, M. H., Jr, Beatty, J. T., Goffeau, A. & 11 other authors (1999b). The major facilitator superfamily. J Mol Microbiol Biotechnol 1, 257–279.

Salomón, R. A. & Farías, R. N. (1995). The peptide antibiotic microcin 25 is imported through the TonB pathway and the SbmA protein.J Bacteriol177, 3323-3325.[Abstract]

Sanchez, J. C., Gimenez, R., Schneider, A., Fessner, W. D., Baldoma, L., Aguilar, J. & Badia, J. (1994). Activation of a cryptic gene encoding a kinase for L-xylulose opens a new pathway for the utilisation of L-lyxose by Escherichia coli.J Biol Chem269, 29665-29669.[Abstract/Free Full Text]

Sanders, J. W., Leenhouts, K., Burghoorn, J., Brands, J. R., Venema, G. & Kok, J. (1998). A chloride-inducible acid resistance mechanism in Lactococcus lactis and its regulation. Mol Microbiol27, 299-310.[Medline]

Saraste, M. & Walker, J. E. (1982). Internal sequence repeats and the path of polypeptide in mitochondrial ADP/ATP translocase.FEBS Lett144, 250-254.[Medline]

Sarker, R. I., Ogawa, W., Shimamoto, T., Shimamoto, T. & Tsuchiya, T. (1997). Primary structure and properties of the Na+/glucose symporter (SglS) of Vibrio parahaemolyticus.J Bacteriol179, 1805-1808.[Abstract]

Sarsero, J. P. & Pittard, A. J. (1995). Membrane topology analysis of Escherichia coli K-12 Mtr permease by alkaline phosphatase and ß-galactosidase fusions.J Bacteriol177, 297-306.[Abstract]

Sarsero, J. P., Wookey, P. J., Gollnick, P., Yanofsky, C. & Pittard, A. J. (1991). A new family of integral membrane proteins involved in transport of aromatic amino acids in Escherichia coli.J Bacteriol173, 3231-3234.[Medline]

Sato, H., Tamba, M., Ishii, T. & Bannai, S. (1999). Cloning and expression of a plasma membrane cystine/glutamate exchange transporter composed of two distinct proteins.J Biol Chem274, 11455-11458.[Abstract/Free Full Text]

Saurin, W., Hofnung, M. & Dassa, E. (1999). Getting in or out: early segregation between importers and exporters in the evolution of ATP-binding cassette (ABC) transporters.J Mol Evol48, 22-41.[Medline]

Schroers, A., Burkovski, A., Wohlrab, H. & Krämer, R. (1998). The phosphate carrier from yeast mitochondria: dimerization is a prerequisite for function.J Biol Chem273, 14269-14276.[Abstract/Free Full Text]

Shao, Z-Q., Lin, R. T. & Newman, E. B. (1994). Sequencing and characterization of the sdaC gene and identification of the sdaCB operon in Escherichia coli K-12. Eur J Biochem222, 901-907.[Abstract]

Shayakul, C., Steel, A. & Hediger, M. A. (1996). Molecular cloning and characterization of the vasopressin-regulated urea transporter of rat kidney collecting ducts.J Clin Invest98, 2580-2587.[Abstract/Free Full Text]

Shukla, V. K. & Chrispeels, M. J. (1998). Aquaporins: their role and regulation in cellular water movement. In NATO-ASI Series, subseries H: Cellular Integration of Signalling Pathways in Plant Development, pp. 11–22. Edited by F. L. Schavio & others. New York: Springer.

Six, S., Andrews, S. C., Unden, G. & Guest, J. R. (1994). Escherichia coli possesses two homologous anaerobic C4-dicarboxylate membrane transporters (DcuA and DcuB) distinct from the aerobic dicarboxylate transport system (Dct).J Bacteriol176, 6470-6478.[Abstract]

Slotboom, D. J., Konings, W. N. & Lolkema, J. S. (1999). Structural features of the glutamate transporter family.Microbiol Mol Biol Rev63, 293-307.[Abstract/Free Full Text]

Sophianopoulou, V. & Diallinas, G. (1995). Amino acid transporters of lower eukaryotes: regulation, structure and topogenesis.FEMS Microbiol Rev16, 53-75.[Medline]

Steiner, H.-Y., Song, W., Zhang, L., Naider, F., Becker, J. M. & Stacey, G. (1994). An Arabidopsis peptide transporter is a member of a new class of membrane transport proteins. Plant Cell6, 1289-1299.[Abstract/Free Full Text]

Steiner, H.-Y., Naider, F. & Becker, J. M. (1995). The PTR family: a new group of peptide transporters.Mol Microbiol16, 825-834.[Medline]

Stephens, R. S., Kalman, S., Lammel, C. J., Fan, R., Marathe, J. & Aravind, L. (1998). Genome sequence of an obligate intracellular pathogen of humans: Chlamydia trachomatis.Science282, 754-759.[Abstract/Free Full Text]

Stucky, K., Hagting, A., Klein, J. R., Matern, H., Henrich, B., Konings, W. N. & Plapp, R. (1995). Cloning and characterization of brnQ, a gene encoding a low-affinity, branched-chain amino acid carrier in Lactobacillus delbruckii subsp. lactis DSM 7290.Mol Gen Genet249, 682-690.[Medline]

Sullivan, T. D., Strelow, L. I., Illingworth, C. A., Phillips, R. L. & Nelson, O. E.Jr (1991). Analysis of the maize brittle-1 alleles and a defective suppressor-mutator induced mutable allele.Plant Cell3, 1337-1348.[Abstract/Free Full Text]

Swift, S., Throup, J. P., Williams, P., Salmond, G. P. C. & Stewart, G. S. A. B. (1996). Quorum sensing: a population-density component in the determination of bacterial phenotype.Trends Biochem Sci21, 214-219.[Medline]

Tauch, A., Hermann, T., Burkovski, A., Krämer, R., Pühler, A. & Kalinowski, J. (1998). Isoleucine uptake in Corynebacterium glutamicum ATCC 13032 is directed by the brnQ gene product.Arch Microbiol169, 303-312.[Medline]

Tsay, Y.-F., Schroeder, J. I., Feldmann, K. A. & Crawford, N. M. (1993). The herbicide sensitivity gene CHL1 of Arabidopsis encodes a nitrate-inducible nitrate transporter.Cell72, 705-713.[Medline]

Turk, E. & Wright, E. M. (1997). Membrane topology motifs in the SGLT cotransporter family.J Membr Biol159, 1-20.[Medline]

Tzagoloff, A., Jang, J., Glerum, D. M. & Wu, M. (1996). FLX1 codes for a carrier protein involved in maintaining a proper balance of flavin nucleotides in yeast mitochondria.J Biol Chem271, 7392-7397.[Abstract/Free Full Text]

Unden, G. & Bongaerts, J. (1997). Alternative respiratory pathways of Escherichia coli: energetics and transcriptional regulation in response to electron acceptors.Biochim Biophys Acta1320, 217-234.[Medline]

Verrey, F., Jack, D. L., Paulsen, I. T., Saier, M. H.Jr & Pfeiffer, R. (1999). New glycoprotein-associated amino acid transporters.J Membr Biol172, 181-192.[Medline]

Vrljic, M., Sahm, H. & Eggeling, L. (1996). A new type of transporter with a new type of cellular function: L-lysine export from Corynebacterium glutamicum.Mol Microbiol22, 815-826.[Medline]

Vrljic, M., Garg, J., Bellman, A. & 7 other authors (1999). The LysE superfamily: topology of the lysine exporter LysE of Corynebacterium glutamicum, a paradigm for a novel superfamily of transmembrane solute translocators. J Mol Microbiol Biotechnol 1, 327–336.[Medline]

Walker, J. E. & Runswick, M. J. (1993). The mitochondrial transport protein superfamily.J Bioenerg Biomembr25, 435-446.[Medline]

Walshaw, D. L. & Poole, P. S. (1996). The general L-amino acid permease of Rhizobium leguminosarum is an ABC uptake system that also influences efflux of solutes. Mol Microbiol21, 1239-1252.[Medline]

Walshaw, D. L., Lowthorpe, S., East, A. & Poole, P. S. (1997). Distribution of a sub-class of bacterial ABC polar amino acid transporter and identification of an N-terminal region involved in solute specificity. FEBS Lett414, 397-401.[Medline]

West, I. C. (1997). Ligand conduction and the gated-pore mechanism of transmembrane transport.Biochim Biophys Acta1331, 213-234.[Medline]

Wookey, P. J. & Pittard, A. J. (1988). DNA sequence of the gene (tyrP) encoding the tyrosine-specific transport system of Escherichia coli.J Bacteriol170, 4946-4949.[Medline]

Yang, B. & Verkman, A. S. (1998). Urea transporter UT3 functions as an efficient water channel.J Bacteriol272, 9369-9372.

Young, G. B., Jack, D. L., Smith, D. W. & Saier, M. H.Jr (1999). The amino acid/auxin:proton symport permease family.Biochim Biophys Acta1415, 306-322.[Medline]

Zakataeva, N. P., Aleshin, V. V., Tokmakova, I. L., Troshin, P. V. & Livshits, V. A. (1999). The novel transmembrane Escherichia coli proteins involved in amino acid efflux.FEBS Lett452, 228-232.[Medline]

Zarbiv, R., Grunewald, M., Kavanaugh, M. P. & Kanner, B. I. (1998). Cysteine scanning of the surroundings of an alkali-ion binding site of the glutamate transporter GLT-1 reveals a conformationally sensitive residue.J Biol Chem273, 14231-14237.[Abstract/Free Full Text]

Zhou, J. J., Theodoulou, F. L., Muldin, I., Ingemarsson, B. & Miller, A. J. (1998). Cloning and functional characterization of a Brassica napus transporter that is able to transport nitrate and histidine.J Biol Chem273, 12017-12023.[Abstract/Free Full Text]