Expression Cloning of Mouse cDNA of
CMP-NeuAc:Lactosylceramide
2,3-Sialyltransferase, an Enzyme That
Initiates the Synthesis of Gangliosides*
Satoshi
Fukumoto
§¶,
Hiroshi
Miyazaki
¶,
George
Goto§,
Takeshi
Urano
,
Keiko
Furukawa
, and
Koichi
Furukawa
From the
Department of Biochemistry II, Nagoya
University School of Medicine, 65 Tsurumai, Nagoya 466-0065 and the
§ Department of Pediatric Dentistry, Nagasaki University
School of Dentistry, 1-7-1, Sakamoto, Nagasaki 852-8102, Japan
 |
ABSTRACT |
Expression cloning of a cDNA for the
2,3-sialyltransferase (GM3 synthase) (EC 2.4.99.-) gene was
performed using a GM3-lacking mouse fibroblast line L cell and anti-GM3
monoclonal antibody. Plasmids from a cDNA library generated with
poly(A)+ RNA of a mouse fibrosarcoma line CMS5j and
pdl3027 (polyoma T antigen) were co-transfected into L cells. The
isolated cDNA clone pM3T-7 predicted a type II membrane protein
with 13 amino acids of cytoplasmic domain, 17 amino acids of
transmembrane region, and a large catalytic domain with 329 amino
acids. Introduction of the cDNA clone into L cells resulted in the
neo-synthesis of GM3 and high activity of
2,3-sialyltransferase.
Among glycosphingolipids, only lactosylceramide showed significant
activity as an acceptor, indicating that this gene product is a
sialyltransferase specific for the synthesis of GM3. An amino acid
sequence deduced from the cloned cDNA showed the typical sialyl
motif with common features among
2,3-sialyltransferases. Among
various mouse tissues, brain, liver, and testis showed relatively high
expression of a 2.3-kilobase mRNA, whereas all tissues, more or
less, expressed this gene.
 |
INTRODUCTION |
Gangliosides are acidic, sialic acid-containing glycosphingolipids
and are ubiquitously expressed in all tissues and cells (1, 2). Their
carbohydrate structures are strictly regulated, depending on the
development of tissues, differentiation, and malignant transformation
of cells (3, 4). Ganglioside synthesis is catalyzed by
glycosyltransferases, and the combination of expression levels of
various glycosyltransferase genes determines characteristic patterns of
ganglioside expression (5-8). Among the gangliosides, GM3 is the
simplest, having only three sugars, and is widely distributed in all
over the body of vertebrates (1). Because GM3 is a precursor for all
other ganglioside series gangliosides and hematosides (7), its
biosynthesis strongly affects the levels of the more complex gangliosides.
Recent progress in the molecular cloning of glycosyl-transferase genes
responsible for the synthesis of gangliosides has enabled us and other
investigators to analyze the regulatory mechanisms for the ganglioside
expression and biological functions of them by manipulation of those
genes to modify the ganglioside profiles in cultured cells and in
experimental animals (9). In our laboratory, we have isolated genes
encoding ganglioside GM2/GD21
synthase (10), GD3 synthase gene (11), and GM1/GD1b/GA1 synthase (12)
and have analyzed expression of these genes in various tissues and
malignant cells (8, 13, 14). We have also established gene knock-out
mice lines in which all complex ganglioside series gangliosides are
genetically eliminated in the whole body (15). The resulting phenotypic
changes in those mutant mice pose new questions about the substitutive
roles of simpler ganglioside structures (e.g. GM3 and GD3)
for the eliminated complex gangliosides.
In the present study, we isolated a mouse cDNA clone that
determines expression of ganglioside GM3 and have analyzed its
expression pattern in various tissues. This gene product corresponds to
GM3-specific sialyltransferase (CMP-NeuAc:lactosylceramide
2,3-sialyltransferase,
2,3S-T) (EC 2.4.99.-). Because of the
critical role that GM3 synthase plays in the biosynthesis of all
gangliosides, this probe will be extremely useful for the analysis of
the biological functions of acidic glycosphingolipids.
 |
MATERIALS AND METHODS |
Tissue Cultures and Cells--
Cell lines were cultured in
Dulbecco's modified Eagle's minimal essential medium supplemented by
10% fetal bovine serum. L cell and B78 (a subline of mouse B16
melanoma) were kindly provided by Dr. A. P. Albino at
Sloan-Kettering Cancer Center in New York. L cell is a mouse fibroblast
line with no
2,3-sialyltransferase activity (for lactosylceramide
acceptor) (16) and minimal GM3 expression. B78 expresses GM3
ganglioside almost exclusively (10).
Plasmids and cDNA Library--
A cDNA library of mouse
fibrosarcoma CMS5j was prepared using poly(A)+ RNA and the
pCDM8 expression vector (Invitrogen, San Diego, CA). This library
contained 5.0 × 106 independent colonies. The strain
of bacterial host used was Escherichia coli MC1061/P3.
Expression Cloning of Mouse GM3 Synthase cDNA--
Plasmids
of the cDNA library were amplified, purified, and then transfected
into L cells together with plasmid pdl3027 (polyoma T gene kindly
provided by C. Basilico (17)) using DEAE-dextran (Amersham Pharmacia
Biotech) as described previously (10). Subconfluent L cells, 2 × 106 in 10-cm dishes (Corning, Corning, NY), were
co-transfected with 8 µg each of cDNA library plasmid and
pdl3027. After 48-60 h, the transfected cells were detached from
plates and incubated with a mouse monoclonal antibody M2590 (mouse IgM)
(18) (Nihon Biotest Research Institute, Tokyo, Japan) at 20 µg/ml on
ice for 45 min. After washing, cells were plated on dishes coated with goat anti-mouse IgM (Cappel, Durham, NC) as described previously (10).
Plasmid DNA was rescued from the panned cells by preparing Hirt
extracts and transformed into MC1061/P3. Expanded plasmids were
transfected again, and the same procedure was repeated five times.
Using isolated colony extracts and microscale DEAE-dextran transfection
and immunofluorescence assay, we isolated a cDNA clone that
determines the expression of ganglioside GM3 in L cells.
DNA Sequencing--
The sequence was determined by
dideoxynucleotide termination sequencing using the PRISM dye terminator
cycle sequencing kit and model 373 DNA sequencer (Applied Biosystems,
Foster City, CA). The cDNA insert of the cloned pM3T-7 was cleaved
by HindIII and NotI and then inserted into
HindIII/NotI sites of phagemid BlueScript
vector. Deletion mutants of this clone were
prepared with a Kilo-Sequence deletion kit (Takara, Kyoto).
Transfection of the Cloned cDNA--
Isolated cDNA clone
pM3T-7 (in pCDM8) was transfected into L cells by the DEAE-dextran
method, and the transfectant cells were used for the analyses of
ganglioside expression and of the enzyme activity after 60 h.
Plasmid pM3T-7/MIKneo was constructed by inserting
XhoI/NotI fragment of pM3T-7 into
XhoI/NotI sites of pMIKneo expression vector
(kindly provided by Dr. K. Maruyama at Tokyo Medical Dental School).
Enzyme Assay--
The enzyme activity of
2,3S-T was measured
as described previously (19). Briefly, to prepare membrane fractions,
samples were lysed using a nitrogen cavitation apparatus. Nuclei were removed by low speed centrifugation, and the supernatant was
centrifuged at 105,000 × g for 1 h at 4 °C. To
analyze enzyme activity of
2,3S-T, the reaction mixture contained
the following in a volume of 50 µl: 100 mM sodium
cacodylate-HCl (pH 6.0), 10 mM MgCl2, 0.3%
Triton CF-54 (Sigma), 325 µM lactosylceramide (Lac-Cer)
(Sigma), 400 mM CMP-NeuAc (Sigma),
CMP-[14C]NeuAc (105 dpm) (NEN Life Science
Products), and membrane containing 30 µg of protein. After incubation
for 3 h at 37 °C in a shaking water bath, the products were
isolated by a C18 Sep-Pak cartridge (Warters, Milford, MA) and analyzed
by TLC and fluorography as described (16). The protein concentration in
the membrane preparation was determined by Lowry's methods (20).
Glycosphingolipids and asialo-fetuin were purchased from Sigma.
RNA Extraction and Northern Blotting--
Total RNA was
extracted from various tissues of mice with acid phenol methods as
described previously (21). 20 µg each of total RNA was applied and
separated on 1.0% agarose/formaldehyde gel. After blotting onto nylon
membrane (GeneScreen Plus, Dupont), they were hybridized with
[32P]dCTP-labeled cDNA probe of pM3T-7. The
expression levels of the gene among tissues were estimated by using a
Bio-Imaging Analyzer BAS2000 (Fuji Film, Tokyo).
Extraction of Gangliosides and TLC Analysis--
Gangliosides
were extracted according to Furukawa et al. (22). Briefly,
glycolipids were extracted from about 400 µl of packed cells of
transfectants and control cells transfected with a vector alone using
chloroform/methanol (2:1, 1:1, and 1:2) sequentially. After desalting,
gangliosides were isolated by DEAE-Sephadex A-50 (Amersham Pharmacia
Biotech) ion exchange chromatography. TLC was performed on a high
performance TLC plates (Merck) using the solvent system
chloroform:methanol:2.5 N NH4OH (60:35:8) and
sprayed by resorcinol for detection of bands. For standard
gangliosides, bovine brain ganglioside mixture (Wako Chemical Inc.,
Tokyo) and GM3 (Snow-brand, Tokyo) were used.
Homology Search--
Nucleotide and amino acid sequence homology
search was carried out using the internet program BLAST (National
Center for Biotechnology Information). Amino acid sequence and
hydropathy analyses were performed with a software GENETYX-MAC version
8.0 (Software Development, Tokyo, Japan).
Flow Cytometry--
Cell surface expression of gangliosides was
analyzed by flow cytometry (Becton Dickinson, Mountain View, CA) using
anti-GM3 monoclonal antibody M2590. The cells were incubated with
monoclonal antibodies for 45 min on ice and stained with fluorescein
isothiocyanate-conjugated goat anti-mouse IgM (Cappel). Intensity of
staining was measured in arbitrary units as the log of fluorescent
intensity and displayed on a 4 decade scale. Control cells for flow
cytometry were prepared by using second antibody alone.
 |
RESULTS |
Isolation of
2,3S-T cDNA Clones--
L cells were
transfected with a cDNA library prepared from CMS5j and pdl3027
containing the polyoma T gene. After five cycles of transfection,
panning, and Hirt extraction, a cDNA clone pM3T-7 was obtained.
Synthesis of GM3 in the Transfectant Cells of the Cloned
cDNA--
pM3T-7 in pCDM8 or pMIKneo vector was transiently
introduced into L cells, which lack GM3 expression and GM3 synthase
activity (16), to confirm in flow cytometry whether the clone
determines the expression of GM3 on the cell surface. As shown in Fig.
1, L cells transfected with pM3T-7
expressed definite amount of GM3, whereas those transfected with the
vector alone did not. Glycosphingolipids extracted from the
transfectant cells were analyzed to confirm GM3 synthesis. As shown in
Fig. 2, the transfectant cells with pM3T-7/MIKneo showed a definite GM3 band in TLC, although the parent
cell (data not shown) and the transfectant cells with pMIKneo alone
showed a very faint band, suggesting that the parent L cell expresses a
minimal level of the enzyme.

View larger version (19K):
[in this window]
[in a new window]
|
Fig. 1.
Transient expression of GM3 in L cells.
A, GM3 synthesis reaction. B, flow cytometry of
cDNA transfectant cells. Cloned cDNA pM3T-7 in pCDM8
(right panel) or pCDM8 alone (left panel) was
transfected by DEAE-dextran into L cells. Expression of GM3 was
analyzed after 60 h using GM3-specific monoclonal antibody M2590
in flow cytometry as described under "Materials and Methods." The
x axis is fluorescence intensity, and the y axis
is relative cell number. Thin lines are with monoclonal
antibody, and thick lines are controls.
|
|

View larger version (24K):
[in this window]
[in a new window]
|
Fig. 2.
Synthesis of GM3 in the L cells transfected
with 2,3S-T cDNA. L cells were
transiently transfected with pM3T-7/MIKneo or pM3T-7/CDM8 (data not
shown), and glycosphingolipids were extracted after 60 h as
described under "Materials and Methods." A, the acidic
fractions were isolated and separated in TLC. St,
ganglioside standard of bovine brain gangliosides (totally 10 µg)
plus purified GM3 (0.3 µg). Lane L cell/pMIKneo is
gangliosides from L cells transfected with pMIKneo alone. Lane L
cell/ 2,3S-T/pMIKneo is gangliosides from L cells transfected
with 1 µg/ml of pM3T-7/MIKneo. The solvent used was chloroform,
methanol, 0.2% CaCl2 (60:35:8). Resorcinol spray was
performed for the detection of bands. B, comparison of GM3
quantity measured by the intensities of standard GM3 bands (data not
shown) and bands in A using a densitometer.
|
|
Sequence of the Insert of pM3T-7--
Fig.
3 shows the entire sequence of the insert
of pM3T-7 determined by sequence analysis of the original pM3T-7 in
pCDM8 or deletion constructs of phagemid BlueScript
containing XhoI/NotI fragment. Total size of this
insert was 1710 base pairs comprising a 12-base pair 5'-untranslated
region, a continuous open reading frame of 1077 base pairs, and 621 base pairs of 3'-untranslated region. The initiation codon at the
beginning of the open reading frame is embedded within a sequence
similar to the Kozak consensus initiation sequence (23, 24). This open
reading frame predicts a 359-amino acid protein with a molecular mass
of 41,244 daltons. Search of currently available protein and nucleic
acid data bases identified a number of genes with significant sequence
homology to this cDNA. A majority of those genes were
sialyltransferases, and highly homologous regions were detected at the
sequences of sialylmotif L, sialyl motif S, and a few other regions.
Among sialyl-transferase cDNAs, ST3Gal III (ST3N)
(Gal
1,3/1,4GlcNAc
2,3S-T) generally showed the highest homology.2 The second group
is ST3Gal III (Gal
1,3GalNAc/Gal
1,4GlcNAc
2,3S-T). The next
groups are ST3Gal IA (ST3O) (Gal
1,3GalNAc
2,3S-T) and ST3Gal II
(ST3GalA2) (Gal
1,3GalNAc
2,3S-T). These latter two groups showed
approximately similar levels of homology. The next groups are ST6
groups and ST8Sia groups, which showed the lowest homology among these
sialyltransferases. Inspection and hydropathy of the predicted protein
sequence suggested that this enzyme molecule has a similar structural
organization to those of reported glycosyltransferase genes (Fig. 3). A
single hydrophobic segment with 17 amino acids was present at near the
amino terminus. This putative signal anchor sequence would place 13 residues within the cytosolic compartment and 329 amino acids within
the Golgi lumen as a catalytic domain. The predicted amino acid
sequence indicated the presence of three N-glycosylation
sites at amino acids 180-182, 224-226, and 334-336 (Fig. 3).

View larger version (56K):
[in this window]
[in a new window]
|
Fig. 3.
Nucleotide sequences of cloned
2,3S-T pM3T-7. Top, deduced amino
acid sequences for the single open reading frame. The putative
transmembrane region with 17 amino acids was double
underlined. Candidates of N-glycosylation site were
marked by a single underline. Bottom, hydropathy
analysis of the coding region based on the deduced amino acids
according to Hopp and Woods (40).
|
|
Products of the
2,3S-T cDNA Can Catalyze Only GM3
Synthesis--
We then analyzed the enzyme activity of GM3 synthase in
membrane extracts from those transfectants. As shown in Fig.
4A, the transfectant cells
with pM3T-7/MIKneo (
2,3S-T/pMIKneo) showed very high GM3
synthase activity when lactosylceramide was used as an acceptor (1394 units). On the other hand, transfectant cells with pMIKneo alone were
completely negative. Thus, this clone was able to determine the
expression of GM3 and
2,3S-T activity when introduced into a mouse
cell line, strongly suggesting that this cDNA encodes the GM3
synthase. Using the membrane extracts, the enzyme activity was examined
toward various acceptor compounds such as galactosylceramide,
glucosylceramide, asialo-GM2 (GA2), asialo-GM1 (GA1), GM3, GM1, GD1b,
GD1a, GT1b, etc., as shown in Fig. 4B. Asialo-fetuin was
also examined. None of the acceptors examined showed significant levels
of [14C]NeuAc incorporation except lactosylceramide,
indicating that this sialyltransferase is almost specific for the
synthesis of GM3 as reported by Iber et al. (25). Only GA1
and GA2 showed minimal levels of incorporation. Asialo-fetuin also
showed no incorporation of [14C]NeuAc (data not
shown).

View larger version (20K):
[in this window]
[in a new window]
|
Fig. 4.
2,3S-T activity in the extracts
from transient transfectant cells of the cloned cDNA.
A, enzyme activity with lactosylceramide as acceptor was
measured as described under "Materials and Methods." Membrane
fractions were prepared from B78 and L cells transfected with pMIKneo
alone or pM3T-7/MIKneo ( 2,3S-T/pMIKneo). As shown in the
inset, GM3 band was observed in the TLC of the products.
B, comparison of enzyme activity for various acceptor
structures (325 µM each was used) as indicated in the
figure.
|
|
Northern Blot Analysis--
Expression levels of the
2,3S-T
gene in various tissues of mice were examined by Northern blots using
total RNA. Among various tissues examined, brain showed the highest
level, and then testis, heart and liver also showed relatively high
levels, whereas almost all tissues showed some levels of the gene
expression (Fig. 5). Mouse melanoma B78
also expressed a high level of transcript, consistent with the
serological data indicating high levels of GM3 expression.

View larger version (41K):
[in this window]
[in a new window]
|
Fig. 5.
Northern blots of
2,3S-T (GM3 synthase) gene. A, 20 µg
each of total RNA from mouse tissues was separated on agarose gel and
then blotted onto nylon membrane. Hybridization with
32P-labeled probe derived from pM3T-7 was performed as
described under "Materials and Methods." B, relative
expression levels of mRNA of 2,3S-T gene among mouse tissues
were measured by Bio-Imaging Analyzer (Fuji, Tokyo) and presented as a
percentage of the value of B78.
|
|
 |
DISCUSSION |
We have identified a cDNA that determines the expression of
GM3 and induces
2,3S-T activity in transfected cells. Furthermore, the deduced amino acid sequence indicated the presence of the sialyl
motifs, which has been recognized among all mammalian and chicken
sialyltransferases cloned so far (26), suggesting that this cDNA
represents the GM3 synthase gene. Examination of the sequence showed
features in common with other
2,3-sialyltransferases. As shown in
Fig. 6, the derived GM3 synthase sequence
contained a number of characteristic regions commonly detected only in
the sequences of ST3Gal
2,3-sialyltransferases (see enclosed
boxes in sialyl motif L and S). These regions may be involved in
the recognition of acceptor structures. The conserved YPE sequence was
also noted by Sasaki in a recent review (27). In addition, the ST8Sia
group showed a common sequence (NPS) in the carboxyl terminus region of
the sialyl motif L. This sequence contrasted with the sequence YPE,
which was commonly detected among ST3Gal group, supporting the idea
that this region might be involved in the acceptor recognition.

View larger version (48K):
[in this window]
[in a new window]
|
Fig. 6.
Sequence alignment of sialyl motif
regions of representative sialyltransferases. Amino acid sequences
of sialyl motif L and S of nine sialyltransferases were aligned.
Top, sialyl motif L. Bottom, sialyl motif S. Identical amino acid(s) in the majority of sialyltransferases are
indicated by bold letters, and identical amino acids only
among ST3Gal(s) were enclosed by boxes. Derivations are as
follows: ST3Gal(GM3), reported in this paper; ST3Gal I, ST3GalA1 (41);
ST3Gal II, ST3Gal A.2 (42); ST3Gal III, mST3Gal III (43); ST3Gal IV,
mST3Gal IV (43); ST6GalNAc II, Gal 1,3GalNAc-specific GalNAc 2,6S-T
(44); ST8SiaI, GD3 synthase (11); ST8SiaII, polysialic acid synthase
(45); ST8SiaIV, N-glycan 2,8S-T (46).
|
|
The properties of the GM3 synthase predicted from the amino acid
sequence (Fig. 3, A and B) and demonstrated by
the analysis with extracts from cDNA-transfected cells were not so
different from those of previously purified GM3 synthases. For example, Km for lactosylceramide and CMP-NeuAc of the GM3
synthase expressed in the transfected cells of pM3T-7 were 270 µM and 313 µM, respectively (data not
shown). These values were of similar magnitude to the results of Preuss
et al. (28), i.e. 80 µM
(Km for lactosylceramide) and 210 µM
(Km for CMP-NeuAc), respectively. High specificity
for lactosyl-ceramide was also similar, although a few acceptors
besides lactosylceramide also showed low levels of activity in their
results. On the other hand, the predicted molecular mass of the enzyme
was 41,244 Da, whereas that in their paper was 76 kDa in
SDS-polyacrylamide gel electrophoresis. This discrepancy might be
partially due to the glycosylation of the enzyme protein. Actually, the
predicted amino acid sequence contained three possible sites of
N-glycosylation (Fig. 3A). Otherwise, differences
of the tissues and species used as sources of mRNA and enzymes need
to be considered.
Among gangliosides, GM3 has the simplest carbohydrate structure and is
located at the starting point of the synthesis of all ganglioside
series gangliosides. Almost all mammalian tissues and cells contain GM3
(1), suggesting its important biological role. Moreover, a number of
studies have shown that GM3 functions in the regulation of
platelet-derived growth factor receptor (29) and epidermal growth
factor receptor (30-32) in cell adhesion (33-35), in the regulation
of keratinocyte proliferation (36), and in the promotion of the
differentiation of HL60 leukemia cells (37). Recently, Yamamura
et al. (38) reported that GM3 co-localized with various
signaling molecules such as c-Src and a low molecular GTP-binding
protein Rho on the cell surface of B16 forming a glycolipid-enriched microdomain. Therefore, GM3 itself might play important roles at
diverse situations for the regulation of cell proliferation and
differentiation. Following the isolation of the GM3 synthase gene, the
effects of GM3 endogenously generated in the cells can be studied to
confirm the possible functions described above. Moreover, human
cDNA of GM3 synthase has recently been cloned by Ishii et
al. (39), enabling us to analyze the regulatory mechanisms of
leukocyte differentiation by gangliosides.
Besides the importance of GM3 itself,
2,3S-T is also important,
because it can regulate the amount of all gangliosides in particular
tissues and cells. For example, although a majority of human melanoma
cells characteristically express ganglioside GD3, they also express
very high level of GM3. The high GD2 expression in neuroblastoma cells
is also partly based on the high expression of GM3. Because GM3
expression is ubiquitous, its importance is often not appreciated.
However, the quantity of GM3 is quite different among different normal
tissues and various tumor cells. GM3 seems to control the expression
levels of more specific and complex gangliosides by adjusting the
amounts of precursor structures. Therefore, the mechanisms for the
regulation of the gene expression of
2,3S-T are of great importance
and interest.
Mammalian brain tissues usually express low levels of GM3 compared with
the level of complex gangliosides. However, the actual synthesis of GM3
should be much higher than expected from the amount of GM3, because all
complex ganglioside series gangliosides are synthesized through GM3.
Results of Northern blotting clearly indicated that GM3 synthesis takes
place most actively in the brain among the normal tissues tested (Fig.
5). During the study of substrate specificity of the cloned
2,3S-T,
we found that GD1b specifically inhibited the synthesis of GM3 with
endogenous acceptor lactosylceramide in membrane preparations (data not
shown). This fact indicates that later members of the biosynthetic
pathway (GD1b in this case) may regulate
2,3S-T activity by a
feedback mechanism in the brain tissue. The regulatory mechanisms of
GM3 expression at the levels of gene expression and post-translation are presently being investigated.
 |
ACKNOWLEDGEMENTS |
We thank Dr. Y. Yoshikai in the Department of
Bioprotection at Nagoya University for providing the opportunity to use
the flow cytometry machine and Dr. K. O. Lloyd at Memorial
Sloan-Kettering Cancer Center for carefully reading the manuscript.
 |
FOOTNOTES |
*
This work was supported by Grants-in-Aid for Scientific
Research of Priority Areas 05274103 and Core of Excellence from the Ministry of Education, Science, Sports, and Culture of Japan.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AB013302.
¶
These authors contributed equally to this work.
To whom correspondence should be addressed: Dept. of
Biochemistry II, Nagoya University School of Medicine, 65 Tsurumai,
Showa-ku, Nagoya 466-0065, Japan. Tel.: 81-52-744-2070; Fax:
81-52-744-2069.
2
We propose here to assign the name of ST3Gal V
to GM3 synthase.
 |
ABBREVIATIONS |
The abbreviations used are:
GM2, GalNAc
1
4
(NeuAc
2
3)Gal
1
4Glc Cer;
GD2, GalNAc
1
4
(NeuAc
2
8 NeuAc
2
3)Gal
1
4Glc-Cer;
GD3, NeuAc
2
8
NeuAc
2
3 Gal
1
4 Glc-Cer;
GM3, NeuAc
2
3Gal
1
4Glc-Cer;
Lac-Cer (CDH), Gal
1
4Glc-Cer;
GA2, GalNAc
1
4Gal
1
4Glc-Cer;
GM1, Gal
1
3GalNAc
1
4 (NeuAc
2
3)Gal
1
4Glc-Cer;
GA1, Gal
1
3GalNAc
1
4Gal
1
4 Glc-Cer;
GD1a, NeuAc
2
3Gal
1
3GalNAc
1
4(NeuAc
2
3)Gal
1
4Glc-Cer;
GD1b, Gal
1
3GalNAc
1
4(NeuAc
2
8NeuAc
2
3)Gal
1
4Glc-Cer;
GT1b, Neu Ac
2
3Gal
1
3GalNAc
1
4(Neu
Ac
2
8NeuAc
2
3)Gal
1
4Glc-Cer (ganglioside nomenclature
is based on that of Svennerholm (47));
2, 3S-T,
2,3-sialyltransferase (the nomenclature of sialyltransferases is
based on that of Tsuji et al. (48)).
 |
REFERENCES |
-
Wiegandt, H.
(ed)
(1985)
Glycolipids, pp. 199-260, Elsevier Science Publishing Co., Inc., New York
-
Kaufmann, B.,
Basu, S.,
and Roseman, S.
(1967)
in
Inborn Disorders of Sphingolipid Metabolism (Anderson, S. M., and Volk, B. W., eds), pp. 193-213, Pergamon, New York
-
Hakomori, S.
(1981)
Annu. Rev. Biochem.
50,
733-764[CrossRef][Medline]
[Order article via Infotrieve]
-
Furukawa, K.,
and Lloyd, K. O.
(1990)
in
Human Melanoma: From Basic Science to Clinical Application (Ferrone, S., ed), pp. 15-30, Springer, Heidelberg, Germany
-
Pohlentz, G.,
Klein, D.,
Schwarzmann, G.,
Schmitz, D.,
and Sandhoff, K.
(1988)
Proc. Natl. Acad. Sci. U. S. A.
85,
7044-7048[Abstract]
-
Iber, H.,
Kaufmann, R.,
Pohlentz, G.,
Schwarzmann, G.,
and Sandhoff, K.
(1989)
FEBS Lett.
248,
18-22[CrossRef][Medline]
[Order article via Infotrieve]
-
Iber, H.,
and Sandhoff, K.
(1989)
FEBS Lett.
254,
124-128[CrossRef][Medline]
[Order article via Infotrieve]
-
Yamashiro, S.,
Ruan, S.,
Furukawa, K.,
Tai, T.,
Lloyd, K. O.,
Shiku, H.,
and Furukawa, K.
(1993)
Cancer Res.
53,
5305-5400
-
Lloyd, K. O.,
and Furukawa, K.
(1998)
Glycoconj. J.
15,
627-636[CrossRef][Medline]
[Order article via Infotrieve]
-
Nagata, Y.,
Yamashiro, S.,
Yodoi, J.,
Lloyd, K. O.,
Shiku, H.,
and Furukawa, K.
(1992)
J. Biol. Chem.
267,
12082-12089[Abstract/Free Full Text]
-
Haraguchi, M.,
Yamashiro, S.,
Yamamoto, A.,
Furukawa, K.,
Takamiya, K.,
Lloyd, K. O.,
Shiku, H.,
and Furukawa, K
(1994)
Proc. Natl. Acad. Sci. U. S. A
91,
10455-10459[Abstract/Free Full Text]
-
Miyazaki, H.,
Fukumoto, S.,
Okada, M.,
Hasegawa, T.,
Furukawa, K.,
and Furukawa, K.
(1997)
J. Biol. Chem.
272,
24794-24799[Abstract/Free Full Text]
-
Yamashiro, S.,
Okada, M.,
Haraguchi, M.,
Furukawa, K.,
Lloyd, K. O.,
Shiku, H.,
and Furukawa, K.
(1995)
Glycoconj. J
12,
894-900[Medline]
[Order article via Infotrieve]
-
Yamamoto, A.,
Haraguchi, M.,
Yamashiro, S.,
Fukumoto, S.,
Furukawa, K.,
Takamiya, K.,
Atsuta, M.,
Shiku, H.,
and Furukawa, K.
(1996)
J. Neurochem.
66,
26-34[Medline]
[Order article via Infotrieve]
-
Takamiya, K.,
Yamamoto, A.,
Furukawa, K.,
Yamashiro, S.,
Shin, M.,
Okada, M.,
Fukumoto, S.,
Haraguchi, M.,
Takeda, N.,
Fujimura, K.,
Sakae, M.,
Kishikawa, M.,
Shiku, H.,
Furukawa, K.,
and Aizawa, S.
(1996)
Proc. Natl. Acad Sci. U. S. A.
93,
10662-10667[Abstract/Free Full Text]
-
Yamashiro, S.,
Haraguchi, M.,
Furukawa, K.,
Takamiya, K.,
Yamamoto, A.,
Nagata, Y.,
Lloyd, K. O.,
Shiku, H.,
and Furukawa, K.
(1995)
J. Biol. Chem.
270,
6149-6155[Abstract/Free Full Text]
-
Dailey, L.,
and Basilico, C.
(1985)
J. Virol.
54,
739-749[Medline]
[Order article via Infotrieve]
-
Wakabayashi, S.,
Saito, T.,
Shinohara, N.,
Okamoto, S.,
Tomioka, H.,
and Taniguchi, M.
(1984)
J. Invest. Dermatol
83,
128-133[Abstract]
-
Ruan, S.,
and Lloyd, K. O.
(1992)
Cancer Res.
52,
5725-5731[Abstract]
-
Lowry, O. H.,
Rosebrough, N. J.,
Farr, A. L.,
and Randall, R. J.
(1951)
J. Biol. Chem.
193,
265-275[Free Full Text]
-
Chomczynski, P.,
and Sacchi, N.
(1987)
Anal. Biochem.
162,
156-159[CrossRef][Medline]
[Order article via Infotrieve]
-
Furukawa, K.,
Clausen, H.,
Hakomori, S.,
Sakamoto, J.,
Look, K.,
Lundblad, A.,
Mattes, M. J.,
and Lloyd, K. O.
(1985)
Biochemistry
24,
7820-7826[Medline]
[Order article via Infotrieve]
-
Kozak, M.
(1986)
Cell
44,
283-292[Medline]
[Order article via Infotrieve]
-
Kozak, M.
(1989)
J. Cell Biol.
108,
229-241[Abstract]
-
Iber, H.,
vanEchten, G.,
and Sandhoff, K.
(1991)
Eur. J. Biochem
195,
115-120[Abstract]
-
Paulson, J. C.,
and Colley, K. J.
(1989)
J. Biol. Chem
264,
17615-17618[Free Full Text]
-
Sasaki, K.
(1996)
Trends Glycosci. Glycotechnol.
8,
195-215
-
Preuss, U.,
Gu, X.,
Gu, T.,
and Yu, R. K.
(1993)
J. Biol. Chem
268,
26273-26278[Abstract/Free Full Text]
-
Bremer, E. G.,
Hakomori, S.,
Bowen-Pope, D. F.,
Raines, E.,
and Ross, R.
(1984)
J. Biol. Chem.
259,
6818-6825[Abstract/Free Full Text]
-
Bremer, E. G.,
Schlessinger, J.,
and Hakomori, S.
(1986)
J. Biol. Chem
261,
2434-2440[Abstract/Free Full Text]
-
Weis, F. M.,
and Davis, R. J.
(1990)
J. Biol. Chem.
265,
12059-12066[Abstract/Free Full Text]
-
Song, W.,
Welti, R.,
Hafner-Strauss, S.,
and Rintoul, D. A.
(1993)
Biochemistry
32,
8602-8607[Medline]
[Order article via Infotrieve]
-
Kojima, N.,
and Hakomori, S.
(1991)
J. Biol. Chem.
266,
17552-17558[Abstract/Free Full Text]
-
Kojima, N.,
Shiota, M.,
Sadahira, Y.,
Handa, K.,
and Hakomori, S.
(1992)
J. Biol. Chem.
267,
17264-17270[Abstract/Free Full Text]
-
Zheng, M.,
Fang, H.,
Tsuruoka, T.,
Tsuji, T.,
Sasaki, T.,
and Hakomori, S.
(1993)
J. Biol. Chem.
268,
2217-2222[Abstract/Free Full Text]
-
Paller, A. S.,
Amsmeier, S. L.,
Alvarez-Franco, M.,
and Bremer, E. G.
(1993)
J. Invest. Dermatol.
100,
841-845[Abstract]
-
Nojiri, H.,
Takaku, F.,
Terui, Y.,
Miura, Y.,
and Saito, M.
(1986)
Proc. Natl. Acad. Sci. U. S. A.
83,
782-786[Abstract]
-
Yamamura, S.,
Handa, K.,
and Hakomori, S.
(1997)
Biochem. Biophys. Res. Commun
236,
218-222[CrossRef][Medline]
[Order article via Infotrieve]
-
Ishii, A.,
Ohta, M.,
Watanabe, Y.,
Sakoe, K.,
Nakamura, M.,
Inokuchi, J.,
Sanai, Y.,
and Saito, M.
(1998)
J. Biol. Chem
273,
31652-31655[Abstract/Free Full Text]
-
Hopp, T. P.,
and Woods, K. R.
(1981)
Proc. Natl. Acad Sci. U. S. A.
78,
3824-3828[Abstract]
-
Lee, Y.-C.,
Kurosawa, N.,
Hamamoto, T.,
Nakaoka, T.,
and Tsuji, S.
(1993)
Eur. J. Biochem.
216,
377-385[Abstract]
-
Lee, Y.-C.,
Kojima, N.,
Wada, E.,
Kurosawa, N.,
Nakaoka, T.,
Hamamoto, T.,
and Tsuji, S.
(1994)
J. Biol. Chem.
269,
10028-10033[Abstract/Free Full Text]
-
Kono, M.,
Ohyama, Y.,
Lee, Y.-C.,
Hamamoto, T.,
Kojima,
and Tsuji, S.
(1996)
Glycobiology
7,
469-479[Abstract]
-
Kurosawa, N.,
Inoue, M.,
Yoshida, Y.,
and Tsuji, S
(1996)
J. Biol. Chem.
271,
15109-15116[Abstract/Free Full Text]
-
Yoshida, Y.,
Kurosawa, N.,
Kanematsu, T.,
Kojima, N.,
and Tsuji, S.
(1996)
J. Biol. Chem.
271,
30167-30173[Abstract/Free Full Text]
-
Yoshida, Y.,
Kojima, N.,
and Tsuji, S.
(1995)
J. Biochem. (Tokyo)
118,
658-664[Abstract]
-
Svennerholm, L.
(1963)
J. Neurochem.
10,
613-623[Medline]
[Order article via Infotrieve]
-
Tsuji, S.,
Datta, A. K.,
and Paulson, J. C.
(1996)
Glycobiology
6,
v-xiv[Medline]
[Order article via Infotrieve]
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.