STRUCTURAL REQUIREMENTS FOR ADHESION OF SOLUBLE RECOMBINANT MURINE VASCULAR CELL-ADHESION MOLECULE-1 TO ALPHA-4-BETA-1

被引:72
作者
RENZ, ME
CHIN, HH
JONES, S
FOX, J
KIM, KJ
PRESTA, LG
FONG, S
机构
[1] GENENTECH INC,DEPT IMMUNOL,S SAN FRANCISCO,CA 94080
[2] GENENTECH INC,DEPT PROT ENGN,S SAN FRANCISCO,CA 94080
[3] GENENTECH INC,CELL BIOL & METAB GRP,S SAN FRANCISCO,CA 94080
[4] GENENTECH INC,HYBRIDOMA GRP,S SAN FRANCISCO,CA 94080
关键词
D O I
10.1083/jcb.125.6.1395
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
This study describes the identification of seven amino acid residues of the vascular cell adhesion molecule (VCAM-1) that influence binding to the alpha 4 beta 1 receptor. Using recombinant murine VCAM-1-IgG, which is bound by both mouse (WEHI 231) and human (Ramos) lymphoid cells, two approaches demonstrated the crucial role of the first two NH2-terminal Ig-like domains in binding: (a) blocking monoclonal anti-mouse VCAM-1 antibodies bound to only truncation variants that included the first two domains; (b) site-direct mutagenesis of the first NH2-terminal domain showed that alanine substitution of the amino acid residues R36, D40, K46, S54, N65, T72, and E81 partially or completely reduced adherence by human and/or mouse cells. Of these D40, when mutated to A, N, or K (but not E), showed complete abrogation of adherence by mouse and human cells, as well as inability to bind blocking antimurine VCAM-1 antibody MVCAM.A429, while not inducing gross structural perturbations in VCAM-1. By molecular modeling, the D40 residue was located on a beta turn connecting two beta strands defined as C and D. The residues R36, K46, S54, N65, T72, and E81, which perturb cell adherence and caused small changes to gross structure, are conformationally near or adjacent to D40. Although these residues, identified as crucial for cell adhesion, are ah located in domain 1, it is evident that there is a structural requirement for domains 1 and 2 to be intact so that cell adhesive function can occur.
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页码:1395 / 1406
页数:12
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