Quaternary organization of GPIb-IX complex and insights into Bernard-Soulier syndrome revealed by the structures of GPIb? and a GPIb?/GPIX chimera.

ABSTRACT: Platelet GPIb-IX receptor complex has 3 subunits GPIb?, GPIb?, and GPIX, which assemble with a ratio of 1:2:1. Dysfunction in surface expression of the complex leads to Bernard-Soulier syndrome. We have crystallized the GPIb? ectodomain (GPIb?(E)) and determined the structure to show a single leucine-rich repeat with N- and C-terminal disulphide-bonded capping regions. The structure of a chimera of GPIb?(E) and 3 loops (a,b,c) taken from the GPIX ectodomain sequence was also determined. The chimera (GPIb?(Eabc)), but not GPIb?(E), forms a tetramer in the crystal, showing a quaternary interface between GPIb? and GPIX. Central to this interface is residue Tyr106 from GPIb?, which inserts into a pocket generated by 2 loops (b,c) from GPIX. Mutagenesis studies confirmed this interface as a valid representation of interactions between GPIb? and GPIX in the full-length complex. Eight GPIb? missense mutations identified from patients with Bernard-Soulier syndrome were examined for changes to GPIb-IX complex surface expression. Two mutations, A108P and P74R, were found to maintain normal secretion/folding of GPIb?(E) but were unable to support GPIX surface expression. The close structural proximity of these mutations to Tyr106 and the GPIb?(E) interface with GPIX indicates they disrupt the quaternary organization of the GPIb-IX complex.

SUBMITTER: McEwan PA 

PROVIDER: S-EPMC3217411 | biostudies-literature | 2011 Nov

REPOSITORIES: biostudies-literature