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Thromb Haemost 1978; 40(01): 144-151
DOI: 10.1055/s-0038-1648644
Original Article
Schattauer GmbH Stuttgart

The Action of Immobilized Thrombin on Factor VIII, Fibrinogen and a Synthetic Tripeptide

J E Brown
The Department of Pathology, University of California San Diego, School of Medicine, La Jolla, California 92093, U.S.A.
,
C L Carton
The Department of Pathology, University of California San Diego, School of Medicine, La Jolla, California 92093, U.S.A.
,
A S Dietz
The Department of Pathology, University of California San Diego, School of Medicine, La Jolla, California 92093, U.S.A.
,
R F Baugh
The Department of Pathology, University of California San Diego, School of Medicine, La Jolla, California 92093, U.S.A.
,
C Hougie
The Department of Pathology, University of California San Diego, School of Medicine, La Jolla, California 92093, U.S.A.
› Author Affiliations
Further Information

Publication History

Received 20 July 1977

Accepted 14 February 1978

Publication Date:
12 July 2018 (online)

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Summary

Bovine thrombin was insolubilized by attachment to cyanogen bromide-activated Sepharose (Sepharose-thrombin) or to activated (Affi-Gel 10) agarose containing a 10 Å long arm (Affi-Gel-thrombin). Coupling in both instances approximated 7,000 units of thrombin per ml packed gel as determined by 125I-thrombin incorporation. The thrombin beads hydrolyzed the synthetic tripeptide Bz-Phe-Val-Arg-pNA (S-2160) at different rates, with the Sepharose-thrombin more active (220 esterase units per ml) than Affi-Gel thrombin (20.4 units per ml). The Km was significantly higher for the insolubilized thrombins (2×10-3M) than uncoupled thrombin (Km = 8×10-5M). The Sepharose- thrombin activated factor VIII significantly more rapidly than Affi-Gel-thrombin. Neither matrix-bound thrombin clotted a fibrinogen solution or liberated significant amounts of fibrinopeptides over 48 hr. This data indicates that a proteolysis of factor VIII, rather than a complex with thrombin, is the method of activation of factor VIII and that factor VIII is more accessible to the action of immobilized thrombin than is fibrinogen.

 

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