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Prepublished online as a Blood First Edition Paper on April 17, 2002; DOI 10.1182/blood-2002-02-0343.
Submitted February 4, 2002
Accepted March 13, 2002
Functional characterization of recombinant Factor V Hong Kong and Cambridge
Bjorn Dahlback*, Eva Norstrom, and Elisabeth Thorelli
University of Lund, Department of Clinical Chemistry, Malmo, Sweden
* Corresponding author; email: bjorn.dahlback{at}klkemi.mas.lu.se.
In factor V (FV) Cambridge (Arg306Thr) and Hong Kong (Arg306Gly), a cleavage site for anticoagulant activated protein C (APC) which is crucial for the inactivation of FVa is lost. While patients carrying FV Hong Kong have a normal APC response, those with FV Cambridge were reported to be APC resistant. To elucidate the molecular characteristics of the two FV mutants, we recreated them in a recombinant system and evaluated their functional properties. The two FV variants yielded identical APC-resistance patterns with APC responses being intermediate of those of wild-type FV and FV Leiden (Arg506Gln), which is known to be associated with the APC-resistance phenotype. In the absence of protein S, APC-mediated FVa inactivation curves obtained with the two variants were identical, resulting in partial FVa inactivation. In the presence of protein S, both FVa variants were almost completely inactivated, which was due to protein S stimulation of the cleavage at Arg679. In a FVIIIa-degradation system, both the two FV variants demonstrated slightly impaired APC-cofactor activity. The ability of APC to cleave at both Arg506 and Arg679 in FVa Cambridge and Hong Kong and the slight decrease in APC-cofactor activity of the two FV variants may explain the low thrombotic risk associated with these Arg306 mutations. In conclusion, we demonstrate that recombinant FV Cambridge and Hong Kong behave identical in in vitro assays and provide a mechanism for the low thrombotic risk associated with these FV mutations.
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