Differential Binding of Two Chicken β-Galactoside-Specific Lectins to Homologous Lymphocyte Subpopulations and Evidence for Inhibitor Activity of the Dimeric Lectin on Stimulated T Cells

Schneller, Maximilian; André, Sabine; Cihak, Josef; Kaltner, Herbert; Merkle, Heidrun; Janrademaker, Geert; Haverkamp, Johan; Thomas‐Oates, Jane; Lösch, U.; Gabius, Hans‐Joachim

doi:10.1006/cimm.1995.0005

Cited by 41 publications

(38 citation statements)

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“…Gel filtration analysis (28) of galectin-1 and galectin-1(L11A) showed no difference (17.64 and 17.59 min, respectively) between the retention times of the two proteins, indicating that the mutation had no detectable impact on dimerization or shape. This was confirmed by hemagglutination assays (37); the minimal lectin concentration required to yield hemagglutination was 0.15 g/ml for both galectin-1 and galectin-1(L11A). The hemagglutination assay also permits determination of carbohydrate-binding capacity in a semiquantitative manner, as assessed by the inhibition of hemagglutination with serial dilutions of the haptenic sugar lactose (38).…”

Section: Structural Convergence Among the Geranylgeranyl-binding Domamentioning

confidence: 56%

Galectin-1(L11A) Predicted from a Computed Galectin-1 Farnesyl-Binding Pocket Selectively Inhibits Ras-GTP

et al. 2004

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Ras biological activity necessitates membrane anchorage that depends on the Ras farnesyl moiety and is strengthened by Ras/galectin-1 interactions. We identified a hydrophobic pocket in galectin-1, analogous to the Cdc42 geranylgeranyl-binding cavity in RhoGDI, possessing homologous isoprenoid-binding residues, including the critical L11, whose RhoGDI L77 homologue changes dramatically on Cdc42 binding. By substituting L11A, we obtained a dominant interfering galectin-1 that possessed normal carbohydrate-binding capacity but inhibited H-Ras GTP-loading and extracellular signal-regulated kinase activation, dislodged H-Ras(G12V) from the cell membrane, and attenuated H-Ras(G12V) fibroblast transformation and PC12-cell neurite outgrowth. Thus, independently of carbohydrate binding, galectin-1 cooperates with Ras, whereas galectin-1(L11A) inhibits it.

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Section: Structural Convergence Among the Geranylgeranyl-binding Domamentioning

confidence: 56%

Galectin-1(L11A) Predicted from a Computed Galectin-1 Farnesyl-Binding Pocket Selectively Inhibits Ras-GTP

et al. 2004

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“…Presented at the cell surface and acting via its carbohydrate binding capacity, galectin-1 (or its avian homologue CG-16) is mitogenic for murine Thy-1-negative spleen or lymph node cells, mammalian vascular and hu- man Hs729 cells, and SK-UT-1 osteosarcoma cells (52)(53)(54)(55)(56). In contrast, overexpression of galectin-1 during glucocorticoid-dependent induction of apoptosis in human T-cell CEM C7 leukemia cells and its carbohydrate-dependent proapoptotic activity on activated T-cells show that the responses to lectin binding are cell type-specific (57)(58)(59)(60). Of potential clinical relevance in this respect is the observation that galectin-1 appears to contribute to the elimination of CD7ϩ T cells during disease progression in Sézary syndrome.…”

Section: Discussionmentioning

confidence: 99%

Negative Regulation of Neuroblastoma Cell Growth by Carbohydrate-dependent Surface Binding of Galectin-1 and Functional Divergence from Galectin-3

Kopitz¹,

Reitzenstein²,

André³

et al. 2001

Journal of Biological Chemistry

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262

192

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The cell density-dependent growth inhibition of human SK-N-MC neuroblastoma cells is initiated by increased ganglioside sialidase activity leading to elevated cell surface presentation of ganglioside GM1, a ligand of galectin-1. We herein show that the extent of the cell surface expression of the galectin coincides with marked increases of the sialidase activity. Reverse transcriptase-polymerase chain reaction analysis excludes a regulation at the transcriptional level. Exposure of cells to purified galectin-1 reveals its carbohydrate-dependent activity to reduce cell proliferation. Assays to detect DNA fragmentation biochemically and cytometrically and to block caspases render it unlikely that galectin-1 acts as a classical proapoptotic factor on these cells. Because the chimeric galectin-3 shares binding sites and binding parameters with galectin-1 for these cells, we tested whether this galectin will elicit the same response as the homodimeric cross-linking galectin-1. Evidently, galectin-3 fails to affect cell growth by itself but interferes with galectin-1 upon coincubation. Its proteolytically truncated variant, the C-terminal lectin domain with impaired capacity to form aggregates when surface bound, has only weak binding properties. Thus, the way in which the galectin-1 interacts topologically with an apparently common set of ligands relative to galectin-3 is crucial for eliciting post-binding events. We conclude that galectin-1 is a probable effector in the sialidase-dependent growth control in this system. Moreover, the experiments with galectin-3 reveal functional divergence, most probably based on different topologies of presentation of homologous carbohydratebinding sites.

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“…The ␤-galactoside-binding lectin from chicken liver was purified by successive application of affinity and ion-exchange chromatography. The homogeneity was ascertained by two-dimensional gel electrophoresis, as described (9,10).…”

Section: Preparation Of the Ligand A Mixture Of 1-o-[3ј-(trifluoroacmentioning

confidence: 99%

NMR-Based, Molecular Dynamics- and Random Walk Molecular Mechanics-Supported Study of Conformational Aspects of a Carbohydrate Ligand (Galβ1-2Galβ1-R) for an Animal Galectin in the Free and in the Bound State

Siebert

Gilleron

Kaltner

et al. 1996

Biochemical and Biophysical Research Communications

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The binding of a carbohydrate to a lectin may affect the conformation of the ligand. To address this question for the galectin from chicken liver, the conformation of Gal␤1-2Gal␤1-R was analyzed in the free and in the galectin-bound state with 2D-ROESY-and 1D-as well as 2D-transferred NOE-experiments. A computerassisted analysis of spatial parameters of the ligand by molecular dynamics (MD) and random walk molecular mechanics (RAMM) calculations, taking different dielectric constants from ⑀ ‫ס‬ 1 to ⑀ ‫ס‬ 80 and various force fields into account, were instrumental to define the energetic minima of the free state. NMR-derived interresidual distance constraints enabled a conformational mapping. The two overlapping interresidual distance constraints obtained from transferred-NOE experiments of the galectin-ligand complex clearly support the notion that the conformation of the disaccharide in the bound state is at least very close to its global energy minimum state in solution. © 1996 Academic Press, Inc.Protein-carbohydrate interactions can guide crucial physiological reactions such as cell adhesion, inter-and intracellular glycoprotein routing and regulation of various cellular functions (1, 2). Their evident importance prompts to analyze in detail the molecular characteristics of this type of recognition in solution. The combination of computer-assisted calculations of the conformation and NMR-based experiments for the free ligand and lectin-saccharide complex provides a suitable tool to address this question. It enables us to compare spatial parameters of a certain ligand before and after association with members of a lectin family with similar binding specificity, e.g. to galactosides. Agglutinins of this class mediate cell-matrix interactions and enhance immune functions (3-5). Due to the remarkable affinity of Gal␤1-2Gal␤1-R to the immunomodulatory Viscum album agglutinin, reported previously (6, 7), we have chosen to map conformational aspects of this high-affinity ligand for galactoside-binding lectins of organisms from different branches of the evolutionary tree. Herein, we report the first study in this area for an animal galectin. Transferred nuclear Overhauser effect (TRNOE)-based experiments in combination with conformational distance mapping, random walk molecular mechanics (RAMM) and molecular dynamics (MD) calculations indicate that the actual conformations of this disaccharide in the complex and in the free form are at least very similar.

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Differential Binding of Two Chicken β-Galactoside-Specific Lectins to Homologous Lymphocyte Subpopulations and Evidence for Inhibitor Activity of the Dimeric Lectin on Stimulated T Cells

Cited by 41 publications

References 0 publications

Galectin-1(L11A) Predicted from a Computed Galectin-1 Farnesyl-Binding Pocket Selectively Inhibits Ras-GTP

Galectin-1(L11A) Predicted from a Computed Galectin-1 Farnesyl-Binding Pocket Selectively Inhibits Ras-GTP

Negative Regulation of Neuroblastoma Cell Growth by Carbohydrate-dependent Surface Binding of Galectin-1 and Functional Divergence from Galectin-3

NMR-Based, Molecular Dynamics- and Random Walk Molecular Mechanics-Supported Study of Conformational Aspects of a Carbohydrate Ligand (Galβ1-2Galβ1-R) for an Animal Galectin in the Free and in the Bound State

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