2 Department of Endocrinology, Kagawa Medical University, Kagawa 761-0793, Japan; 3 GalPharma Co., Ltd., Kagawa 761-0301, Japan; 4 Research Equipment Center, Kagawa Medical University, Kagawa 761-0793, Japan; and 5 Department of Immunology and Immunopathology, Kagawa Medical University, Kagawa 761-0793, Japan
Received on March 13, 2003; revised on July 7, 2003; accepted on July 10, 2003
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Abstract |
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Key words: galectin / inflammation / integrin / matrix metalloproteinase / neutrophil
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Introduction |
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Currently, 10 members of the human galectin family (galectins-14, galectins-710, galectins-12 and -13) are known. These family members can be classified into three subtypes according to their structures (Hirabayashi and Kasai, 1993): the prototype (galectins-1, -2, -7, -10, and -13) and chimera-type (galectin-3) galectins have a single carbohydrate recognition domain (CRD), and they form a homodimer resulting in homobifunctional cross-linking activity. Galectins-4, -8, -9, and -12 belong to the tandem repeat type, consisting of two CRDs joined by a linker peptide. N-terminal and C-terminal CRDs of the tandem repeat type galectins generally have different sugar binding specificities (Wasano and Hirakawa, 1999
; Arata et al., 2001
; Sato et al., 2002
; Hirabayashi et al., 2002
), and this heterobifunctional property makes it possible to cross-link a wide variety and combination of glycoconjugates. Due to this multiple cross-linking potential, tandem repeat-type galectins are attractive targets for studying novel function of the galectin family in the immune system.
Galectin-8 was first identified in a rat liver cDNA library as the third member (mouse but not human galectin-6 is known) of the murine tandem repeat type galectin family (Hadari et al., 1995). Two independent studies concerning tumor-associated antigens resulted in the identification of human galectin-8 cDNAs (Su et al., 1996
; Bassen et al., 1999
). In contrast to other tandem repeat type galectins, galectin-8 is widely expressed in normal tissues in addition to tumor cells. The physiological role of galectin-8 is largely unknown. However, it seems that galectin-8 is involved in malignant transformation and cellmatrix interaction (Danguy et al., 2001
; Hadari et al., 2000
; Levy et al., 2001
).
In our search for novel function of galectins in the immune system, especially in the innate immune system, it was found that galectin-8 induced firm and reversible adhesion of peripheral blood neutrophils in vitro. Neutrophils play a central role in innate immunity to bacterial infection. Recruitment of circulating neutrophils to an affected site proceeds through several defined steps, namely, attachment to, rolling on, and firm adhesion to endothelial cells and then transendothelial migration. Recent studies have revealed the molecular basis of the leukocyteendothelium interaction. More than a dozen molecules, on both neutrophil and endothelial cell membranes, involved in the interaction have been identified. Here, we report the characterization of galectin-8 as a novel modulator of neutrophil function.
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Results |
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Identification of galectin-8 receptors in neutrophils
To examine the molecular mechanisms by which galectin-8 induces neutrophil adhesion, proteins bound to galectin-8 in a lactose-sensitive manner were affinity-purified from the solubilized neutrophil membrane fraction. Only two major protein bands corresponding to molecular weights of about 180,000 and 94,000 were detected with sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDSPAGE) (Figure 4A). The N-terminal amino acid sequences of the higher- and lower-molecular-weight band materials were determined to be FNLDTENAMTFQENAR and APRQRQSTLXLFP, respectively. These sequences completely matched with those of integrin M/CD11b and promatrix metalloproteinase-9 (proMMP-9). A mutant galectin-8 with an inactivated N-terminal CRD (galectin-8R69H) exhibited similar affinity for integrin
M and reduced affinity for proMMP-9, compared to the wild type (Figure 4A). On the other hand, another mutant form with an inactivated C-terminal CRD (galectin-8R233H) retained affinity for proMMP-9 but did not bind integrin
M.
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The neutrophil adhesion assay showed that galectin-8R69H possessed reduced but apparent adhesion-inducing activity and that inactivation of C-terminal CRD almost completely abolished this activity (Figure 5). In addition, anti-integrin M antibodies but not anti-integrin
L ones strongly inhibited the neutrophil adhesion induced by galectin-8 (Table I).
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Discussion |
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Integrin M exists as a heterodimer with ß subunit (integrin ß2/CD18). However, we could not detect integrin ß2 in the affinity-purified preparations by N-terminal amino acid sequence analysis and western blot analysis. The loss of integrin ß2 indicates the possibility that the heterodimer structure of Mac-1 changes when galectin-8 interacts with the oligosaccharide moiety of
subunit. The structural change may result in lowered interaction between the two subunits, and thus, ß2 subunit was lost during affinity purification. These results suggest that galectin-8 promoted cell adhesion by activating integrin
M (
Mß2/Mac-1), that is, by inducing a direct interaction between Mac-1 and the substratum, in at least about 60% of the adhering neutrophils. There is, however, the possibility that neutrophil cell surface protein(s) other than integrin
M play a role in galectin-8-induced neutrophil adhesion because galectin-8R69H did not retain full adhesion-inducing activity. Dong and Hughes (1997)
reported that integrin
M is one of the predominant binding proteins for galectin-3 in macrophages. In addition, preliminary experiments showed that human galectin-7 could bind integrin
6 (unpublished data). These results and those of galectin-8 suggest that integrin
subunits are common targets of the members of the galectin family.
The expression of Mac-1 is not restricted to neutrophils; eosinophils and monocytes/macrophages also express the molecule. Currently, the reason for the inability of galectin-8 to induce eosinophil adhesion is unclear. One possible explanation for this observation is that the number of Mac-1 molecules capable of interacting with galectin-8 on eosinophils may be significantly smaller than that on neutrophils. Structural heterogeneity in the oligosaccharide moiety of Mac-1 may also result in the difference.
Kuwabara and Liu (1996) reported that galectin-3 promoted neutrophil adhesion to a laminin-coated plastic surface. In our case, galectins-1 and -3 induced apparent neutrophil adhesion at high concentrations, but the adhering cells could be detached by pipetting, showing that the binding between neutrophils and the substratum induced by galectin-8 is much stronger than that induced by galectins-1 and -3.
ICAM-1, C3bi, collagen, and fibrinogen are known as physiological ligands for Mac-1. Among them ICAM-1 plays a critical role in neutrophil adhesion to endothelial cells and subsequent migration (Diamond et al., 1990). Galectin-8 induced neutrophil adhesion to ICAM-1-coated plates in a manner very similar to that to untreated plates (Figure 2). Because plastic culture plates serve as good substratum in neutrophil adhesion, it is likely that the maximum effect of galectin-8 on neutrophil adhesion was achieved using untreated plates.
MMPs are considered to play a key role in normal tissue remodeling and in pathological destruction of the matrix in various diseases. MMP-9 is produced mainly by polymorphonuclear leukocytes and monocytes/macrophages. For example, transmigrating neutrophils secrete MMP-9 to degrade matrix proteins (Baggiolini and Dewald, 1984). The activities of MMPs are controlled not only through their gene expression but also through processing of their precursor forms and inhibition by endogenous inhibitors. Stromelysins, including MMPs-3 and -10, are capable of activating proMMPs (proMMPs-1, -8 and -9) and are considered to play a pivotal role in the initiation of the MMP activation cascade. In this study, we showed that galectin-8 accelerated the processing of proMMP-9 mediated by MMP-3 using purified components and the experimental conditions that permit slow processing of proMMP-9 in the absence of galectin-8. The N-terminal amino acid sequence analysis revealed that MMP-9 processed under the present conditions (1 µg/ml proMMP-9; proMMP-9:MMP-3 = 1:0.1, molar ratio) was not the fully processed form but possessed 35 extra N-terminal amino acid residues compared to the fully processed one. On the other hand, accelerated production of the fully processed MMP-9 in the presence of galectin-8 was observed in some experiments in which higher concentrations of proMMP-9 and MMP-3 (2.5 µg/ml proMMP-9; proMMP-9:MMP-3 = 1:1, molar ratio) were used (data not shown). It was difficult, however, to obtain reproducible results under the latter conditions. These results suggest that galectin-8 facilitates the processing by recruiting proMMP-9 and MMP-3 (see later discussion) when these proteins exist at low concentrations and that the production of fully processed MMP-9 is sensitive to the experimental conditions when nanomolar concentrations of the MMPs were used. The lactose-sensitive nature of the accelerating effect indicates that interaction between galectin-8 and oligosaccharide chain(s) of proMMP-9 (Rudd et al., 1999
; Mattu et al., 2000
) and/or MMP-3 is indispensable. Although the molecular mechanisms involved in the effect remain to be determined, the inability of monovalent galectin-8 mutants, galectin-8R69H and galectin-8R233H, to accelerate the reaction suggests that the formation of a ternary complex from proMMP-9, galectin-8, and MMP-3 is an essential step. The finding that galectin-8 can bind purified MMP-3 is consistent with this speculation.
Galectin-8 was as effective as fMLP, a potent leukocyte chemotactic factor, in inducing superoxide production by peripheral blood neutrophils. fMLP-induced superoxide production continued only for 2 min in the absence of cytochalasin B. In contrast, galectin-8 showed a long-lasting effect. The difference is probably due to rapid down-regulation of the fMLP receptor and not the galectin-8 receptor after activation. The experiments involving galectin-8 mutants suggested that Mac-1 is a major signal transducer in galectin-8-induced superoxide production. Yamaoka et al. (1995) reported that human galectin-3 stimulated superoxide production by neutrophils, although the maximum activity of galectin-3 was about 25% of that of fMLP. They suggested that the aggregation of NCA-160 (CD66) induced by galectin-3 might be important for neutrophil activation. However, because galectin-3 is capable of interacting with integrin
M, it is possible that galectin-3 triggers superoxide production via interaction with Mac-1.
Neutrophil adhesion to endothelial cells and subsequent transendothelial migration proceed through defined steps: (1) circulating cells are captured and induced to roll on the stimulated endothelium; (2) surface-bound activating signal(s) cause activation of neutrophil ß2 integrins, which immobilize the cells on the endothelium; and (3) cells migrate through the endothelial cell layer and interstitial space by degrading the extracellular matrix. Selectins and ß2 integrins play central roles in the first and second steps, respectively. MMPs and ß2 integrins are important factors in the third step. In addition to these factors, the present study suggests that galectin-8 is a new player in the process of neutrophil migration: galectin-8 may participate in the second and third steps as an activator of Mac-1 and a modulator of MMP-9 activity, respectively. This possibility must be evaluated by means of in vivo studies.
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Materials and methods |
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Site-directed mutagenesis of galectin-8
The following forward and reverse primers were used to generate cDNAs for site-directed mutants of galectin-8: G8R69H-F (5'-CATTTCAATCCTCATTTCAAAAGG-3'), G8R69H-R (5'-CCTTTTGAAATGAGGATTGAAATG-3'), G8R233H-F (5'-CACTTGAACCCACACCTGAATATT-3'), and G8R233H-R (5'-AATATTCAGGTGTGGGTTCAAGTG-3'). Site-directed mutagenesis of galectin-8 residue Arg69 (and Arg233) to His was carried out as described elsewhere (Matsushita et al., 2000). Galectin-8 cDNA carrying a double mutation, R69H and R233H (galectin-8R69, 233H), was prepared by introducing the R233H mutation into galectin-8R69H. The resultant mutations were confirmed by DNA sequencing.
Expression, purification, and sugar-binding activity of recombinant proteins
Expression of recombinant proteins in Escherichia coli BL21 cells was carried out as described previously (Matsushita et al., 2000). Recombinant proteins were purified by affinity chromatography on a lactose-agarose column (Seikagaku, Tokyo) (galectin-8 and galectin-8R233H) or a glutathione-Sepharose column (Amersham Biosciences) (galectin-8R69H and galectin-8R69, 233H). The protein concentration was determined using BCA protein assay reagent (Pierce, Rockford, IL) and bovine serum albumin as a standard. The sugar-binding ability of recombinant galectin-8 was assessed by asialofetuin-agarose affinity chromatography. Purified GST-fusion proteins (1 mg) were applied to an asialofetuin-agarose column (bed volume, 0.5 ml; 20 mg of conjugated protein/ml) equilibrated with phosphate buffered saline (PBS). After washing the column with PBS, proteins bound to the column were eluted with PBS, 0.2 M lactose. A control sample, nonbound fraction, and bound fraction were analyzed by SDSPAGE.
Preparation of antibodies to galectin-8
The affinity-purified GST-galectin-8 was digested with thrombin and the released GST moiety was removed by glutathione-Sepharose affinity chromatography. Anti-galectin-8 antiserum was raised in Japanese white rabbits as described (Shoji et al., 2002).
In vitro cell adhesion assay
Neutrophils and eosinophils were isolated as described previously (Matsumoto et al., 1998; Matsushita et al., 2000
). Isolated cells were added to 24-well tissue culture plates (2.5 x 105 cells in 0.45 ml of medium/well) in triplicate. After the addition of 50 µl of the assay sample, the cells were allowed to adhere for 60 min at 37°C. At the end of the incubation period, loosely attached cells were removed by vigorous pipetting and washing with PBS. The attached cells were recovered by treatment with 0.25% trypsin/0.5 mM ethylenediamine tetra-acetic acid (EDTA) for 10 min at 37°C, and then sonicated for 15 s in 50 mM sodium phosphate buffer (pH 7.4) containing 2 M NaCl and 2 mM EDTA. The DNA content of the sonicate was determined by the method of Labarca and Paigen (1980)
using calf thymus DNA as a standard. In some experiments, tissue-culture plates were pretreated with GST-galectin-8 or recombinant human ICAM-1 (extracellular domain; Genzyme-Techne, Minneapolis, MN) dissolved in PBS for 3 h at room temperature. The plates were washed three times with PBS before the adhesion assay.
Fluorescence microscopy of the neutrophil cytoskeleton and galectin-8
Fluorescein isothiocyanate (FITC)-phalloidin was used to stain F-actin. Neutrophils adhering to a plastic chamber slide (Nalge Nunc International, Naperville, IL) were washed with PBS and then fixed with 4% paraformaldehyde. The cells were permeabilized with 0.1% Triton X-100 in PBS prior to incubation with 1.65 x 10-7 M FITC- phalloidin (Molecular Probes, Eugene, OR) for 20 min, followed by washing with PBS. All samples were covered with 50% glycerol and a coverslip and then subjected to confocal laser scanning microscopy. Immunofluorescent localization of galectin-8 was carried out using affinity-purified anti-galectin-8 polyclonal antibodies. Paraformaldehyde-fixed permeabilized neutrophils were incubated with anti-galectin-8 antibodies (5 µg/ml) overnight at 4°C. After washing with PBS, the cells were incubated with anti-rabbit IgG-FITC (10 µg/ml; Kirkegaard & Perry Laboratories, Gaithersburg, MD) for 1 h at room temperature. The cells were washed with PBS and then subjected to confocal microscopy as described.
Western blot analysis
Samples were electrophoretically separated in SDS/10% polyacrylamide gels and then transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA). Immunodetection was carried out with monoclonal antibodies against human MMP-3 (Daiichi Fine Chemical, Toyama, Japan) and human CD18 (Cymbus Biotechnology, Chandlers Ford, UK) and polyclonal antibodies against human MMP-9 (Chemicon, Temecula, CA) as described previously (Nishi et al., 1995).
Isolation of galectin-8-interacting proteins and protein sequencing
Purified neutrophils suspended in 10 mM TrisHCl (pH 7.5), 1 mM phenylmethylsulfonyl fluoride were subjected to a single freezethaw cycle and then sonicated. The total membrane fraction was isolated by centrifugation at 50,000 x g for 30 min and then suspended in 10 mM TrisHCl (pH 7.5), 0.5 M NaCl, 1.5% Triton X-100 (8 ml/1 x 108 cells). After incubation for 1 h at 4°C, the solubilized membrane fraction was obtained by centrifugation. The resulting supernatant was incubated with recombinant proteins (1 µM) for 1 h at 4°C. After the addition of glutathione-Sepharose gel, the mixture was incubated for 45 min at 4°C with gentle mixing. The gel was washed with 20 mM TrisHCl (pH 7.5), 0.15 M NaCl, 0.03% 3-{(3-cholamidopropyl)dimethylammonio}-1-propane sulfonate (TBS, 0.03% CHAPS). Galectin-binding proteins were eluted with TBS, 0.2 M lactose. The proteins were resolved by means of 10% SDSPAGE and stained with Coomassie brilliant blue R-250 (CBB R-250). To determine the N-terminal amino acid sequences of the galectin-8-interacting proteins, the resolved proteins were transferred to PVDF membranes. The blotted proteins were stained with CBB R-250 and cut out, and then the N-terminal amino acid sequence was determined with a gas-phase sequencer.
Processing of ProMMP-9 with MMP-3
Activated MMP-3 was prepared by treating proMMP-3 (human synovial fibroblasts; Biogenesis, Kingston, NH) with 2 mM aminophenyl mercuric acetate (APMA; Sigma, St. Louis, MO). APMA was removed from the activated MMP-3 preparation by dialysis. The reaction mixture for the processing of proMMP-9 was mad up of 20 mM TrisHCl (pH 7.5), 0.4 M NaCl, 5 mM CaCl2, 1 µM ZnCl2, 0.03% CHAPS, 100 ng proMMP-9 (human neutrophils; Calbiochem, San Diego, CA), and 0.252 µM recombinant protein in a final volume of 100 µl. After the addition of activated MMP-3 (5 ng/100 µl), the reaction mixture was incubated for 060 min at 37°C. Aliquots of the mixture were withdrawn as specified. Activation of proMMP-9 was assessed by western blot analysis.
Determination of the binding activity of galectin-8 with MMPs
Purified MMPs (1 µg) were mixed with GST-galectin-8 (25 µg) in 0.4 ml 20 mM TrisHCl (pH 7.5), 0.4 M NaCl, 5 mM CaCl2, 1 µM ZnCl2, 0.1% Triton X-100. After incubation for 1 h at 4°C, glutathione-Sepharose gel was added to the mixture, followed by further incubation for 45 min at 4°C with gentle mixing. The gel was washed with TBS, 0.03% CHAPS. Galectin-binding proteins were eluted with TBS, 0.2 M lactose. A control sample, nonbound fraction, and bound fraction were subjected to western blot analysis. Activated MMP-9 was prepared by treating proMMP-9 with APMA.
Determination of superoxide production
O2- production was determined by the method of Yamaoka et al. (1995). Purified neutrophils (2.5 x 106/2 ml), suspended in PBS containing 0.5 mM MgCl2, 0.8 mM CaCl2, and 7.5 mM glucose were placed in a cuvette. The cuvette also contained horse heart cytochrome c (75 µM). The reaction was carried out in the presence or absence of cytochalasin B (5 µg/ml; Biomol Research Laboratories, Plymouth Meeting, PA). In control experiments, superoxide dismutase (SOD, 50 µg/ml) was added. After preincubation for 5 min at 37°C, the stimulants were added and the absorbance change at 550 nm was monitored. The O2--generating activity was calculated using a molar extinction coefficient of 20.5 x 103 M-1cm-1.
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Acknowledgements |
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Footnotes |
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Abbreviations |
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References |
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Avni, O., Pur, Z., Yefenof, E., and Baniyash, M. (1998) Complement receptor 3 of macrophages is associated with galectin-1-like protein. J. Immunol., 160, 61516158.
Baggiolini, M. and Dewald, B. (1984) Exocytosis by neutrophils. Contemp. Top. Immunobiol., 14, 221246.[Medline]
Bassen, R., Brichory, F., Caulet-Maugendre, S., Bidon, N., Delaval, P., Desrues, B., and Dazord, L. (1999) Expression of Po66-CBP, a type-8 galectin, in different healthy, tumoral and peritumoral tissues. Anticancer Res., 19, 54295433.[ISI][Medline]
Bidon, N., Brichory, F., Bourguet, P., Le Pennec, J.P., and Dazord, L. (2001) Galectin-8: a complex sub-family of galectins. Int. J. Mol. Med., 8, 245250.[ISI][Medline]
Danguy, A., Rorive, S., Decaestecker, C., Bronckart, Y., Kaltner, H., Hadari, Y.R., Goren, R., Zich, Y., Petein, M., Salmon, I., and others. (2001) Immunohistochemical profile of galectin-8 expression in benign and malignant tumors of epithelial, mesenchymatous and adipous origins, and of the nervous system. Histol. Histopathol., 16, 861868.[ISI][Medline]
Diamond, M.S., Staunton, D.E., de Fougerolles, A.R., Stacker, S.A., Garcia-Aguilar, J., Hibbs, M.L., and Springer, T.A. (1990) ICAM-1 (CD54): a counter-receptor for Mac-1 (CD11b/CD18). J. Cell Biol., 111, 31293139.[Abstract]
Dong, S. and Hughes, R.C. (1997) Macrophage surface glycoproteins binding to galectin-3 (Mac-2-antigen). Glycoconj. J., 14, 267274.[CrossRef][ISI][Medline]
Hadari, Y.R., Arbel-Goren, R., Levy, Y., Amsterdam, A., Alon, R., Zakut, R., and Zick, Y. (2000) Galectin-8 binding to integrins inhibits cell adhesion and induces apoptosis. J. Cell Sci., 113, 23852397.
Hadari, Y.R., Paz, K., Dekel, R., Mestrovic, T., Accili, D., and Zick, Y. (1995) Galectin-8. A new rat lectin, related to galectin-4. J. Biol. Chem., 270, 34473453.
Hirabayashi, J. and Kasai, K. (1993) The family of metazoan metal-independent beta-galactoside-binding lectins: structure, function and molecular evolution. Glycobiology, 3, 297304.[Abstract]
Hirabayashi, J., Hashidate, T., Arata, Y., Nishi, N., Nakamura, T., Hirashima, M., Urashima, T., Oka, T., Futai, M., Muller, W., and others. (2002) Oligosaccharide specificity of galectins: a search by frontal affinity chromatography. Biochim. Biophys. Acta, 1572, 232254.[ISI][Medline]
Kuwabara, I. and Liu, F.-T. (1996) Galectin-3 promotes adhesion of human neutrophils to laminin. J. Immunol., 156, 39393944.[Abstract]
Labarca, C. and Paigen, K. (1980) A simple, rapid, and sensitive DNA assay procedure. Anal. Biochem., 102, 344352.[ISI][Medline]
Levy, Y., Arbel-Goren, R., Hadari, Y.R., Eshhar, S., Ronen, D., Elhanany, E., Geiger, B., and Zick, Y. (2001) Galectin-8 functions as a matricellular modulator of cell adhesion. J. Biol. Chem., 276, 3128531295.
Lowe, J.B. (2001) Glycosylation, immunity, and autoimmunity. Cell, 104, 809812.[ISI][Medline]
Matsumoto, R., Matsumoto, H., Seki, M., Hata, M., Asano, Y., Kanegasaki, S., Stevens, R.L. and Hirashima, M. (1998) Human ecalectin, a variant of human galectin-9, is a novel eosinophil chemoattractant produced by T lymphocytes. J. Biol. Chem., 273, 1697616984.
Matsushita, N., Nishi, N., Seki, M., Matsumoto, R., Kuwabara, I., Liu, F.T., Hata, Y., Nakamura, T., and Hirashima, M. (2000) Requirement of divalent galactoside-binding activity of ecalectin/galectin-9 for eosinophil chemoattraction. J. Biol. Chem., 275, 83558360.
Mattu, T.S., Royle, L., Langridge, J., Wormald, M.R., Van den Steen, P.E., Van Damme, J., Opdenakker, G., Harvey, D.J., Dwek, R.A., and Rudd, P.M. (2000) O-glycan analysis of natural human neutrophil gelatinase B using a combination of normal phase-HPLC and online tandem mass spectrometry: implications for the domain organization of the enzyme. Biochemistry, 39, 1569515704.[CrossRef][ISI][Medline]
Nishi, N., Inui, M., Miyanaka, H., Oya, H., and Wada, F. (1995) Western blot analysis of epidermal growth factor using gelatin-coated polyvinylidene difluoride membranes. Anal. Biochem., 227, 401402.[CrossRef][ISI][Medline]
Ogata, Y., Enghild, J.J., and Nagase, H. (1992) Matrix metalloproteinase 3 (stromelysin) activates the precursor for the human matrix metalloproteinase 9. J. Biol. Chem., 267, 35813584.
Perillo, N.L., Pace, K.E., Seilhamer, J.J., and Baum, L.G. (1995) Apoptosis of T cells mediated by galectin-1. Nature, 378, 736739.[CrossRef][ISI][Medline]
Rabinovich, G.A., Baum, L.G., Tinari, N., Paganelli, R., Natoli, C., Liu, F.-T., and Iacobelli, S. (2002) Galectins and their ligands: amplifiers, silencers or tuners of the inflammatory response? Trends Immunol., 23, 313320.[CrossRef][ISI][Medline]
Rudd, P.M., Mattu, T.S., Masure, S., Bratt, T., Van den Steen, P.E., Wormald, M.R., Kuster, B., Harvey, D.J., Borregaard, N., Van Damme, J., and others. (1999) Glycosylation of natural human neutrophil gelatinase B and neutrophil gelatinase B-associated lipocalin. Biochemistry, 38, 1393713950.[CrossRef][ISI][Medline]
Sano, H., Hsu, D.K., Yu, L., Apgar, J.R., Kuwabara, I., Yamanaka, T., Hirashima, M., and Liu, F.-T. (2000) Human galectin-3 is a novel chemoattractant for monocytes and macrophages. J. Immunol., 165, 21562164.
Sato, M., Nishi, N., Shoji, H., Seki, M., Hashidate, T., Hirabayashi, J., Kasai, K., Hata, Y., Suzuki, S., Hirashima, M., and Nakamura, T. (2002) Functional analysis of the carbohydrate recognition domains and a linker peptide of galectin-9 as to eosinophil chemoattractant activity. Glycobiology, 12, 191197.
Shoji, H., Nishi, N., Hirashima, M., and Nakamura, T. (2002) Purification and cDNA cloning of Xenopus liver galectins and their expression. Glycobiology, 12, 163172.
Su, Z.Z., Lin, J., Shen, R., Fisher, P.E., Goldstein, N.I., and Fisher, P.B. (1996) Surface-epitope masking and expression cloning identifies the human prostate carcinoma tumor antigen gene PCTA-1 a member of the galectin gene family. Proc. Natl Acad. Sci. USA, 93, 72527257.
Vasta, G.R., Quesenberry, M., Ahmed, H., and O'Leary, N. (1999) C-type lectins and galectins mediate innate and adaptive immune functions: their roles in the complement activation pathway. Dev. Comp. Immunol., 23, 401420.[CrossRef][ISI][Medline]
Wada, J., Ota, K., Kumar, A., Wallner, E.I., and Kanwar, Y.S. (1997) Developmental regulation, expression, and apoptotic potential of galectin-9, a beta-galactoside binding lectin. J. Clin. Invest., 99, 24522461.
Wasano, K. and Hirakawa, Y. (1999) Two domains of rat galectin-4 bind to distinct structures of the intercellular borders of colorectal epithelia. J. Histochem. Cytochem., 47, 7582.
Yamaoka, A., Kuwabara, I., Frigeri, L.G., and Liu, F.-T. (1995) A human lectin, galectin-3 (epsilon bp/Mac-2), stimulates superoxide production by neutrophils. J. Immunol., 154, 34793487.
Yang, R.Y., Hsu, D.K., and Liu, F.-T. (1996) Expression of galectin-3 modulates T-cell growth and apoptosis. Proc. Natl Acad. Sci. USA, 93, 67376742.