RESUMEN

Coagulation factor VIII is a glycosylated, non-covalent heterodimer consisting of a heavy chain (A1-A2-B domains) and a light chain (A3-C1-C2 domains). The association of the chains, and the stability and function of the dimer depend on the presence of metal ions. We applied X-ray fluorescence, X-ray crystallographic structure determination with anomalous signals at different wavelengths, and colorimetric measurements to evaluate the metal binding sites in a recombinant factor VIII molecule, turoctocog alfa. We identified a metal binding site in domain A3 dominated by Cu(+) binding and a site in domain A1 dominated by Zn(2+) binding.

Asunto(s)

Factor VIII/química , Factor VIII/metabolismo , Metales/metabolismo , Sitios de Unión , Calcio/química , Calcio/metabolismo , Colorimetría , Cobre/química , Cobre/metabolismo , Metales/química , Modelos Moleculares , Unión Proteica , Espectrometría por Rayos X , Zinc/química , Zinc/metabolismo