Molecular Cloning, Characterization, and Expression of a Human 14-kDa Lectin

Couraud, Pierre Olivier; Casentini-Borocz, D; Bringman, Timothy S.; Griffith, J E; McGrogan, Michael; Nedwin, Glenn E.

doi:10.1016/s0021-9258(19)85087-1

Cited by 76 publications

(18 citation statements)

References 30 publications

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“…b-Sheet hydrogen bonding across the monomer surface in the interface and formation of a hydrophobic core stabilized the contact (Liao et al, 1994;Varela et al, 1999). Because the amino-acid sequence of human galectin-1 (Hirabayashi and Kasai, 1988;Couraud et al, 1989;Hirabayashi et al, 1989;Bladier et al, 1991) shows a high identity (86%, pairwise alignments) to that of bovine galectin-1 , the available crystal structure of bovine galectin-1 (PDB entry 1SLT) was confidently employed to evaluate the scattering data using the program CRYSOL. Also, homology modeling with calculation of surface accessibilities for selected amino acid residues (i.e., His, Tyr, and Trp) and the experimental determination of these parameters by the laser photo-CIDNP (chemically induced dynamic nuclear polarization) technique for human and bovine galectin-1 underscored the validity of this approach (Siebert et al, 1997).…”

Section: Resultsmentioning

confidence: 99%

Detection of Ligand- and Solvent-Induced Shape Alterations of Cell-Growth-Regulatory Human Lectin Galectin-1 in Solution by Small Angle Neutron and X-Ray Scattering

André

Siebert

et al. 2003

Biophysical Journal

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The bioactivity of galectin-1 in cell growth regulation and adhesion prompted us to answer the questions whether ligand presence and a shift to an aprotic solvent typical for bioaffinity chromatography might alter the shape of the homodimeric human lectin in solution. We used small angle neutron and synchrotron x-ray scattering studies for this purpose. Upon ligand accommodation, the radius of gyration of human galectin-1 decreased from 19.1 +/- 0.1 A in the absence of ligand to 18.2 +/- 0.1 A. In the aprotic solvent dimethyl sulfoxide, which did not impair binding capacity, galectin-1 formed dimers of a dimer, yielding tetramers with a cylindrical shape. Intriguingly, no dissociation into subunits occurred. In parallel, NMR monitoring was performed. The spectral resolution was in accord with these data. In contrast to the properties of the human protein, a nonhomologous agglutinin from mistletoe sharing galactose specificity was subject to a reduction in the radius of gyration from approximately 62 A in water to 48.7 A in dimethyl sulfoxide. Evidently, the solvent caused opposite responses in the two tested galactoside-binding lectins with different folding patterns. We have hereby proven that ligand presence and an aprotic solvent significantly affect the shape of galectin-1 in solution.

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Section: Resultsmentioning

confidence: 99%

Detection of Ligand- and Solvent-Induced Shape Alterations of Cell-Growth-Regulatory Human Lectin Galectin-1 in Solution by Small Angle Neutron and X-Ray Scattering

André

Siebert

et al. 2003

Biophysical Journal

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“…The murine mAb MFIM4 (LFA-lce, CDlla) was obtained from Amac Inc. (Westbrook, ME). The anti-galectin-1 polyclonal rabbit antiserum was obtained as described in (21). Galectin-1 cDNA and purified, recombinant galectin-1 expressed in Escherichia coli were obtained as described in (21).…”

Section: Methodsmentioning

confidence: 99%

“…The anti-galectin-1 polyclonal rabbit antiserum was obtained as described in (21). Galectin-1 cDNA and purified, recombinant galectin-1 expressed in Escherichia coli were obtained as described in (21). Biotinylated galectin-1 for flow cytometry experiments was prepared by coupling N-hydroxysuccinimide-biotin (Bethesda Research Laboratories, Gaithersburg, MD) to recombinant galectin-1, according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Human thymic epithelial cells express an endogenous lectin, galectin-1, which binds to core 2 O-glycans on thymocytes and T lymphoblastoid cells.

Baum¹,

Pang²,

Perillo³

et al. 1995

The Journal of Experimental Medicine

238

189

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SummaryThymic epithelial ceils play a crucial role in the selection of developing thymocytes. Thymocyteepithelial cell interactions involve a number of adhesion molecules, including members of the integrin and immunoglobulin superfamilies. We found that human thymic epithelial cells synthesize an endogenous lectin, galectin-1, which binds to oligosaccharide ligands on the surface of thymocytes and T lymphoblastoid cells. Binding of T lymphoblastoid cells to thymic epithelial cells was inhibited by antibody to galectin-1 on the epithelial cells, and by two antibodies, T305 and 2Bll, that recognize carbohydrate epitopes on the T cell surface glycoproteins CD43 and CD45, respectively. T lymphoblastoid ceils and thymocytes bound recombinant galectin-1, as demonstrated by flow cytometric analysis, and lectin binding was completely inhibited in the presence of lactose. The degree of galectin-1 binding to thymocytes correlated with the maturation stage of the cells, as immature thymocytes bound more galectin-1 than did mature thymocytes. Preferential binding of galectin-1 to immature thymocytes may result from regulated expression of preferred oligosaccharide ligands on those cells, since we found that the epitope recognized by the T305 antibody, the core 2 O-glycan structure on CD43, was expressed on cortical, but not medullary cells. The level of expression of the UDP-GIcNAc:Gal31,3GalNAc-R 31, 6GlcNAc transferase (core 2 31, 6 GlcNAc transferase, or C2GnT), which creates the core 20-glycan structure, correlated with the glycosylation change between cortical and medullary cells. Expression of mRNA encoding the C2GnT was high in subcapsular and cortical thymocytes and low in medullary thymocytes, as demonstrated by in situ hybridization. These results suggest that galectin-1 participates in thymocyte--thymic epithelial cell interactions, and that this interaction may be regulated by expression of relevant oligosaccharide ligands on the thymocyte cell surface.

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“…Gal-1 rGal-1 was expressed according to Couraud et al (1989) in Escherichia coli strain BL21DE3 transformed with the plasmid that encodes Gal-1 and pT7IML-1 (a generous gift form Dr. Linda G. Baum, University of California, Los Angeles, School of Medicine) and purified as previously described (Perillo et al, 1997;Iglesias et al, 1998b). To maintain maximal lectin activity, a reducing agent (0.2 mM dithiothreitol) was added.…”

Section: Preparation Of Recombinant Humanmentioning

confidence: 99%

Galectin-1: biphasic growth regulation of Leydig tumor cells

Biron

Iglesias²,

Troncoso³

et al. 2006

Glycobiology

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Galectin-1 (Gal-1) is a widely expressed beta-galactoside-binding protein that exerts pleiotropic biological functions. To gain insight into the potential role of Gal-1 as a novel modulator of Leydig cells, we investigated its effect on the growth and death of MA-10 tumor Leydig cells. In this study, we identified cytoplasmic Gal-1 expression in these tumor cells by cytofluorometry. DNA fragmentation, caspase-3, -8, and -9 activation, loss of mitochondrial membrane potential (DeltaPsim), cytochrome c (Cyt c) release, and FasL expression suggested that relatively high concentrations of exogenously added recombinant Gal-1 (rGal-1) induced apoptosis by the mitochondrial and death receptor pathways. These pathways were independently activated, as the presence of the inhibitor of caspase-8 or -9 only partially prevented Gal-1-effect. On the contrary, low concentrations of Gal-1 significantly promoted cell proliferation, without inducing cell death. Importantly, the presence of the disaccharide lactose prevented Gal-1 effects, suggesting the involvement of the carbohydrate recognition domain (CRD). This study provides strong evidence that Gal-1 is a novel biphasic regulator of Leydig tumor cell number, suggesting a novel role for Gal-1 in the reproductive physiopathology.

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Molecular Cloning, Characterization, and Expression of a Human 14-kDa Lectin

Cited by 76 publications

References 30 publications

Detection of Ligand- and Solvent-Induced Shape Alterations of Cell-Growth-Regulatory Human Lectin Galectin-1 in Solution by Small Angle Neutron and X-Ray Scattering

Detection of Ligand- and Solvent-Induced Shape Alterations of Cell-Growth-Regulatory Human Lectin Galectin-1 in Solution by Small Angle Neutron and X-Ray Scattering

Human thymic epithelial cells express an endogenous lectin, galectin-1, which binds to core 2 O-glycans on thymocytes and T lymphoblastoid cells.

Galectin-1: biphasic growth regulation of Leydig tumor cells

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