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Characterization of the sialic acid-binding site in sialoadhesin by site-directed mutagenesis.
J Biol Chem. 1996 Apr 19; 271(16):9267-72.JB

Abstract

The sialoadhesins are a distinct subgroup of the immunoglobulin superfamily, comprising sialoadhesin, CD22, the myelin-associated glycoprotein, and CD33. They can all mediate sialic acid-dependent binding to cells with distinct specificities. Sialoadhesin is a murine macrophage-restricted cell-surface molecule with 17 extracellular immunoglobulin-like domains that recognizes NeuAc alpha 2-3Gal in N- and O-glycans and interacts preferentially with cells of the granulocytic lineage. Its sialic acid-binding site is located within the NH2-terminal (membrane-distal) V-set domain. Here we have carried out site-directed mutagenesis in an attempt to identify the binding site of sialoadhesin. A subset of nonconservative mutations disrupted sialic acid-dependent binding without affecting binding of three monoclonal antibodies directed to two distinct epitopes of sialoadhesin. A CD8 alpha-based molecular model predicts that these residues form a contiguous binding site on the GFCC'C" beta-sheet of the V-set domain centered around an arginine in the F strand. A conservative mutation of this arginine to lysine also abolished binding. This amino acid is conserved among all members of the sialoadhesin family and is therefore likely to be a key residue in mediating sialic acid-dependent binding of sialoadhesins to cells.

Authors+Show Affiliations

Imperial Cancer Research Fund Laboratories, University of Oxford, United Kingdom.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

8621587

Citation

Vinson, M, et al. "Characterization of the Sialic Acid-binding Site in Sialoadhesin By Site-directed Mutagenesis." The Journal of Biological Chemistry, vol. 271, no. 16, 1996, pp. 9267-72.
Vinson M, van der Merwe PA, Kelm S, et al. Characterization of the sialic acid-binding site in sialoadhesin by site-directed mutagenesis. J Biol Chem. 1996;271(16):9267-72.
Vinson, M., van der Merwe, P. A., Kelm, S., May, A., Jones, E. Y., & Crocker, P. R. (1996). Characterization of the sialic acid-binding site in sialoadhesin by site-directed mutagenesis. The Journal of Biological Chemistry, 271(16), 9267-72.
Vinson M, et al. Characterization of the Sialic Acid-binding Site in Sialoadhesin By Site-directed Mutagenesis. J Biol Chem. 1996 Apr 19;271(16):9267-72. PubMed PMID: 8621587.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of the sialic acid-binding site in sialoadhesin by site-directed mutagenesis. AU - Vinson,M, AU - van der Merwe,P A, AU - Kelm,S, AU - May,A, AU - Jones,E Y, AU - Crocker,P R, PY - 1996/4/19/pubmed PY - 1996/4/19/medline PY - 1996/4/19/entrez SP - 9267 EP - 72 JF - The Journal of biological chemistry JO - J Biol Chem VL - 271 IS - 16 N2 - The sialoadhesins are a distinct subgroup of the immunoglobulin superfamily, comprising sialoadhesin, CD22, the myelin-associated glycoprotein, and CD33. They can all mediate sialic acid-dependent binding to cells with distinct specificities. Sialoadhesin is a murine macrophage-restricted cell-surface molecule with 17 extracellular immunoglobulin-like domains that recognizes NeuAc alpha 2-3Gal in N- and O-glycans and interacts preferentially with cells of the granulocytic lineage. Its sialic acid-binding site is located within the NH2-terminal (membrane-distal) V-set domain. Here we have carried out site-directed mutagenesis in an attempt to identify the binding site of sialoadhesin. A subset of nonconservative mutations disrupted sialic acid-dependent binding without affecting binding of three monoclonal antibodies directed to two distinct epitopes of sialoadhesin. A CD8 alpha-based molecular model predicts that these residues form a contiguous binding site on the GFCC'C" beta-sheet of the V-set domain centered around an arginine in the F strand. A conservative mutation of this arginine to lysine also abolished binding. This amino acid is conserved among all members of the sialoadhesin family and is therefore likely to be a key residue in mediating sialic acid-dependent binding of sialoadhesins to cells. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/8621587/Characterization_of_the_sialic_acid_binding_site_in_sialoadhesin_by_site_directed_mutagenesis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9258(17)31331-5 DB - PRIME DP - Unbound Medicine ER -