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J Pharm Sci. 2005 Jun;94(6):1288-99.

Lipid binding region (2303-2332) is involved in aggregation of recombinant human FVIII (rFVIII).

Author information

1
Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Amherst, NY 14260, USA.

Abstract

Factor VIII (FVIII) is a multi-domain protein that is important in the clotting cascade. Its deficiency causes Hemophilia A, a bleeding disorder. The unfolding of protein domains can lead to physical instability such as aggregation, and hinder their use in replacement therapy. It has been shown that the aggregation of rFVIIII is initiated by small fluctuations in the protein's tertiary structure (Grillo et al., 2001, Biochemistry 40:586-595). We have investigated the domain(s) involved in the initiation of aggregation using circular dichroism (CD), size exclusion chromatography (SEC), fluorescence anisotropy, domain specific antibody binding, and clotting activity studies. The studies indicated that aggregation may be initiated as a result of conformational change in the C2 domain encompassing the lipid-binding region (2303-2332). The presence of O-phospho-L-Serine (OPLS), which binds to the lipid-binding region of FVIII, prevented aggregation of the protein.

PMID:
15858858
PMCID:
PMC2583467
DOI:
10.1002/jps.20340
[Indexed for MEDLINE]
Free PMC Article

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