title: Evaluation of the metal binding sites in a recombinant coagulation factor VIII identifies two sites with unique metal binding properties.
abstract: 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.
authors: Novo Nordisk A/S, Novo Nordisk Park, DK-2760 Måløv, Denmark. AnSv@novonordisk.com
authors: Lars Anders
authors: LA
authors: Svensson
authors: Lars
authors: L
authors: Thim
authors: Ole Hvilsted
authors: OH
authors: Olsen
authors: Else Marie
authors: EM
authors: Nicolaisen
mesh_terms: D001665:Binding Sites; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D003124:Colorimetry; D003300:Copper / Q000737:chemistry / Q000378:metabolism; D005169:Factor VIII / Q000737:chemistry* / Q000378:metabolism*; D008670:Metals / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D011485:Protein Binding; D013052:Spectrometry, X-Ray Emission; D015032:Zinc / Q000737:chemistry / Q000378:metabolism
chemical_list: D008670:Metals; C577506:recombinant factor VIII N8; D003300:Copper; D005169:Factor VIII; D015032:Zinc; D002118:Calcium