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Selectivity of von Willebrand factor triplet bands towards heparin binding supports structural model
Author(s) -
Fischer Bernhard E.,
Thomas Kathy B.,
Schlokat U.,
Dorner F.
Publication year - 1999
Publication title -
european journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0902-4441
DOI - 10.1111/j.1600-0609.1999.tb01740.x
Subject(s) - chemistry , von willebrand factor , recombinant dna , protein subunit , heparin , microbiology and biotechnology , affinity chromatography , binding site , biochemistry , platelet , biology , immunology , enzyme , gene
Human plasma‐derived von Willebrand factor (hp‐vWF) and recombinant von Willebrand factor (r‐vWF) have been fractionated by heparin affinity chromatography followed by multimer analysis using SDS‐agarose gel electrophoresis. Because heparin binding sites are contained in each vWF subunit, high molecular weight multimers of r‐vWF and hp‐vWF, respectively, were eluted with higher salt concentration, in comparison to r‐vWF and hp‐vWF molecules with a low degree of multimerization. Heparin affinity chromatography did not affect the multimer composition of r‐vWF. By contrast, faster migrating satellite bands and slower migrating satellite bands of hp‐vWF exhibited reduced and increased heparin affinity, respectively, compared to the intermediate band of the same triplet. Because heparin binding sites are localised in the N ‐terminal domain of the hp‐vWF subunit, this result confirms a structural model of hp‐vWF (Fischer et al. , Biochem. J. 1998; 331 :483–488) suggested recently, in which the slower migrating satellite bands have excess of one N ‐terminal fragment and the faster migrating satellite bands lack one N ‐terminal fragment, respectively, in comparison with the corresponding intermediate triplet band.