Structure of an integrin-ligand complex deduced from solution x-ray scattering and site-directed mutagenesis

被引:88
作者
Mould, AP
Symonds, EJH
Buckley, PA
Grossmann, JG
McEwan, PA
Barton, SJ
Askari, JA
Craig, SE
Bella, J
Humphries, MJ
机构
[1] Univ Manchester, Wellcome Trust Ctr Cell Matrix Res, Sch Biol Sci, Manchester M13 9PT, Lancs, England
[2] CLRC, Daresbury Lab, Synchrotron Radiat Dept, Warrington WA4 4AD, Cheshire, England
关键词
D O I
10.1074/jbc.M304627200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The structural basis of the interaction of integrin heterodimers with their physiological ligands is poorly understood. We have used solution x-ray scattering to visualize the head region of integrin alpha(5)beta(1) in an inactive (Ca2+-occupied) state, and in complex with a fragment of fibronectin containing the RGD and synergy recognition sequences. Shape reconstructions of the data have been interpreted in terms of appropriate molecular models. The scattering data suggest that the head region undergoes no gross conformational changes upon ligand binding but do lend support to a proposed outward movement of the hybrid domain in the beta subunit. Fibronectin is observed to bind across the top of the head region, which contains an alpha subunit beta-propeller and a beta subunit vWF type A domain. The model of the complex indicates that the synergy region binds on the side of the beta-propeller domain. In support of this suggestion, mutagenesis of a prominent loop region on the side of the propeller identifies two residues (Tyr(208) and Ile(210)) involved in recognition of the synergy region. Our data provide the first view of a complex between an integrin and a macromolecular ligand in solution, at a nominal resolution of similar to10 Angstrom.
引用
收藏
页码:39993 / 39999
页数:7
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