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- 저자 : 이창규(Suhng Wook Kim) (검색결과 1 건)
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Mutation of individual protein C cleavage sites in Human coagulation factor Va
김성욱(Suhng Wook Kim),이승관(Suhng Wook Kim),이창규(Suhng Wook Kim) 고려대학교 보건과학연구소 2004 보건과학논집 Vol.30 No.1
Resistance to activated protein C (APC) is the most common inherited risk factor for venous thrombosis. In the present study we have investigated the effect of mutations of the APC cleavage sites at Arg<SUB>306</SUB>, Arg<SUB>506</SUB> and Arg<SUB>679</SUB> in the factor V heavy chain on the inactivation of factor Va by APC. Mutants R306A, R506Q and R679A were expressed using B-domain deleted factor V constructs in COS-7 cells. The specific activity of the purified mutant proteins was identical to the wild-type protein (rHFVa) and APC cleavage at the mutated sites was blocked. Characterization of these mutants using a clinical assay for APC resistance demonstrated that R506Q was resistant to APC, but that R306A and R679A were sensitive. In clotting assays, R506Q showed delayed inactivation by APC, whereas R306A showed a rapid but incomplete loss of activity. APC inactivation of R679A was rapid and indistinguishable from rHFVa. These results indicate that mutation of individual APC cleavage sites in factor Va leads to variable degrees of APC. These findings have significant implications for the identification of patients with APC resistance and the understanding of the importance of individual APC cleavage sites in regulating the activity of the prothrombinase complex.
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