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Localization of the putative sialic acid-binding site on the immunoglobulin superfamily cell-surface molecule CD22.
J Biol Chem. 1996 Apr 19; 271(16):9273-80.JB

Abstract

B-lymphocyte antigen CD22 is a member of the recently described sialoadhesin family of immunoglobulin-like cell-surface glycoproteins that bind glycoconjugates terminating in sialic acid. One prominent ligand for CD22 is the highly glycosylated leukocyte surface protein CD45. Using surface plasmon resonance spectroscopy, we characterized the interaction of recombinant mouse CD22 with native CD45 purified from rat thymus (CD45-thy). By in situ desialylation and resialylation of immobilized CD45-thy, we show that mouse CD22 binds to the sialoglycoconjugate NeuGc alpha 2-6Gal beta 1-4GlcNAc carried on CD45-thy N-glycans. Previous studies have shown that the sialic acid-binding site lies within the two membrane-distal domains of CD22 (domains 1 and 2), which are V-set and C2-set immunoglobulin superfamily domains, respectively. To further localize the binding site, we have made 42 single amino acid substitutions throughout both domains. All 12 mutations that abrogated binding to CD45-thy without disrupting antibody binding were of residues within the GFCC'C" beta-sheet of domain 1. These residues are predicted to form a contiguous binding site centered around an arginine residue in the F strand that is conserved in all members of the sialoadhesin family. Our results provide further evidence that immunoglobulin superfamily cell adhesion molecules use the GFCC'C" beta-sheet of membrane-distal V-set domains to bind structurally diverse ligands, suggesting that this surface is favored for cell-cell recognition.

Authors+Show Affiliations

Medical Research Council Cellular Immunology Unit, Sir William Dunn School of Pathology, 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

8621588

Citation

van der Merwe, P A., et al. "Localization of the Putative Sialic Acid-binding Site On the Immunoglobulin Superfamily Cell-surface Molecule CD22." The Journal of Biological Chemistry, vol. 271, no. 16, 1996, pp. 9273-80.
van der Merwe PA, Crocker PR, Vinson M, et al. Localization of the putative sialic acid-binding site on the immunoglobulin superfamily cell-surface molecule CD22. J Biol Chem. 1996;271(16):9273-80.
van der Merwe, P. A., Crocker, P. R., Vinson, M., Barclay, A. N., Schauer, R., & Kelm, S. (1996). Localization of the putative sialic acid-binding site on the immunoglobulin superfamily cell-surface molecule CD22. The Journal of Biological Chemistry, 271(16), 9273-80.
van der Merwe PA, et al. Localization of the Putative Sialic Acid-binding Site On the Immunoglobulin Superfamily Cell-surface Molecule CD22. J Biol Chem. 1996 Apr 19;271(16):9273-80. PubMed PMID: 8621588.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Localization of the putative sialic acid-binding site on the immunoglobulin superfamily cell-surface molecule CD22. AU - van der Merwe,P A, AU - Crocker,P R, AU - Vinson,M, AU - Barclay,A N, AU - Schauer,R, AU - Kelm,S, PY - 1996/4/19/pubmed PY - 1996/4/19/medline PY - 1996/4/19/entrez SP - 9273 EP - 80 JF - The Journal of biological chemistry JO - J Biol Chem VL - 271 IS - 16 N2 - B-lymphocyte antigen CD22 is a member of the recently described sialoadhesin family of immunoglobulin-like cell-surface glycoproteins that bind glycoconjugates terminating in sialic acid. One prominent ligand for CD22 is the highly glycosylated leukocyte surface protein CD45. Using surface plasmon resonance spectroscopy, we characterized the interaction of recombinant mouse CD22 with native CD45 purified from rat thymus (CD45-thy). By in situ desialylation and resialylation of immobilized CD45-thy, we show that mouse CD22 binds to the sialoglycoconjugate NeuGc alpha 2-6Gal beta 1-4GlcNAc carried on CD45-thy N-glycans. Previous studies have shown that the sialic acid-binding site lies within the two membrane-distal domains of CD22 (domains 1 and 2), which are V-set and C2-set immunoglobulin superfamily domains, respectively. To further localize the binding site, we have made 42 single amino acid substitutions throughout both domains. All 12 mutations that abrogated binding to CD45-thy without disrupting antibody binding were of residues within the GFCC'C" beta-sheet of domain 1. These residues are predicted to form a contiguous binding site centered around an arginine residue in the F strand that is conserved in all members of the sialoadhesin family. Our results provide further evidence that immunoglobulin superfamily cell adhesion molecules use the GFCC'C" beta-sheet of membrane-distal V-set domains to bind structurally diverse ligands, suggesting that this surface is favored for cell-cell recognition. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/8621588/Localization_of_the_putative_sialic_acid_binding_site_on_the_immunoglobulin_superfamily_cell_surface_molecule_CD22_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0021-9258(17)31332-7 DB - PRIME DP - Unbound Medicine ER -