Abstract
Sialoadhesin and CD22 are members of a recently characterized family of sialic acid-dependent adhesion molecules belonging to the immunoglobulin superfamily. Sialoadhesin is a macrophage-restricted receptor containing 17 extracellular Ig-like domains which recognizes oligosaccharides terminating in NeuAc alpha 2-3Gal in N- and O-linked glycans. CD22 is a B cell-restricted receptor with seven Ig-like domains which selectively recognizes oligosaccharides terminating in NeuAc alpha 2-6Gal in N-glycans. Sequence similarity between these proteins is highest within their first four amino-terminal Ig-like domains. Here we identify the domain(s) containing the binding sites of both molecules by generating a series of extracellular domain deletion mutants fused to the Fc portion of human IgG1. Binding activity was analyzed by solid phase cell adhesion assays and also by surface plasmon resonance using purified glycophorin and CD45 as ligands for sialoadhesin and CD22, respectively. For sialoadhesin, the amino-terminal V-set Ig-like domain was both necessary and sufficient to mediate sialic acid-dependent adhesion of the correct specificity. In contrast, for murine CD22, only constructs containing both the V-set domain and the adjacent C2-set domain were able to mediate sialic acid-dependent binding. These results are consistent with the sialic acid binding site for both proteins residing in the membrane distal V-set domain, but for CD22 a direct contribution in binding from the neighboring C2-set domain cannot be excluded.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal
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Antigens, CD / chemistry*
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Antigens, CD / metabolism
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Antigens, Differentiation, B-Lymphocyte / chemistry*
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Antigens, Differentiation, B-Lymphocyte / metabolism
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Base Sequence
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Binding Sites
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Carbohydrate Sequence
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Cell Adhesion
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Cell Adhesion Molecules / chemistry
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Cell Adhesion Molecules / metabolism
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DNA Primers
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Epitopes / analysis
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Humans
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Immunoglobulin Fc Fragments / biosynthesis
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Immunoglobulin Fc Fragments / chemistry
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Immunoglobulin G / biosynthesis
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Immunoglobulin G / chemistry
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Immunoglobulins / chemistry*
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Lectins*
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Membrane Glycoproteins / biosynthesis
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Membrane Glycoproteins / chemistry*
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Membrane Glycoproteins / metabolism*
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Mice
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Molecular Sequence Data
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Polymerase Chain Reaction
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Rats / immunology
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Receptors, Immunologic / biosynthesis
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Receptors, Immunologic / chemistry*
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Receptors, Immunologic / metabolism*
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / metabolism
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Sequence Homology, Amino Acid
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Sialic Acid Binding Ig-like Lectin 1
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Sialic Acid Binding Ig-like Lectin 2
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Sialic Acids / metabolism*
Substances
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Antibodies, Monoclonal
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Antigens, CD
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Antigens, Differentiation, B-Lymphocyte
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CD22 protein, human
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Cd22 protein, mouse
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Cell Adhesion Molecules
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DNA Primers
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Epitopes
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Immunoglobulin Fc Fragments
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Immunoglobulin G
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Immunoglobulins
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Lectins
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Membrane Glycoproteins
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Receptors, Immunologic
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Recombinant Fusion Proteins
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SIGLEC1 protein, human
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Sialic Acid Binding Ig-like Lectin 1
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Sialic Acid Binding Ig-like Lectin 2
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Sialic Acids
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Siglec1 protein, mouse