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Thromb Haemost 2009; 102(04): 623-633
DOI: 10.1160/TH09-04-0238
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Differences in prethrombin-1 activation with human or bovine factor Va can be attributed to the heavy chain

Paul Y. Kim
1   Department of Biochemistry, Queen’s University, Kingston, Ontario, Canada
,
Reginald Manuel
1   Department of Biochemistry, Queen’s University, Kingston, Ontario, Canada
,
Michael E. Nesheim
1   Department of Biochemistry, Queen’s University, Kingston, Ontario, Canada
2   Department of Medicine, Queen’s University, Kingston, Ontario, Canada
› InstitutsangabenFinancial support: This work was supported by grant MT-9781 from the Canadian Institutes of Health Research (to MEN) and by grant HS4441 from the Heart and Stroke Foundation of Canada (to MEN). This work was also supported by a Heart and Stroke Foundation of Canada Doctoral Research Award (to PYK).
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Publikationsverlauf

Received: 09. April 2009

Accepted after major revision: 02. Juli 2009

Publikationsdatum:
24. November 2017 (online)

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Summary

Human and bovine factor Va (FVa) function similarly in the activation of prothrombin but differently in the activation of prethrombin-1 (Pre-1). Pre-1 activation with human FVa proceeds at about 22 percent of the rate with bovine FVa. The dependencies of initial rates on the FVa and Pre-1 concentrations indicate that the differential activity is expressed in kcat differences, rather than differences in the assembly of prothrombinase or the Km value of the substrate. The heavy and light chains of both species of FVa were separated and interspecies hybrids were constructed in the presence of Ca++. Studies of the activation of Pre-1 with these hybrids indicate that the species difference can be attributed specifically to the heavy chain of FVa. Analyses of the reactions by SDS-PAGE indicated that cleavage at Arg271 occurs at about the same rate with both species of FVa, but cleavage at Arg320 with human FVa is specifically retarded. A major difference in primary structure between the human and bovine FVa heavy chains comprises 10 residues at COOH-terminus, adjacent to the negatively charged hirudin-like DYDYQ sequence. These residues have pI values of 12.5 and 4.26 in human and bovine FVa, respectively. The lower value would complement the negatively charged DYDYQ sequence but the higher value would counteract it. Thus, we suggest that the differences in the COOH-terminus of the heavy chain are responsible for the differences in Pre-1 activation, and that it specifically influences cleavage at Arg320 in Pre-1.

Keywords

Coagulation factors - factor Va - prethrombin-1 - prothrombin - activation
 

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