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Thromb Haemost 1978; 39(01): 193-200
DOI: 10.1055/s-0038-1646669
Original Article
Schattauer GmbH Stuttgart

On the Preparation of Bovine α-Thrombin

Erwin F Workman Jr
The Departments of Pathology and Biochemistry and the Dental Research Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27514, U.S.A.
,
Roger L Lundblad
The Departments of Pathology and Biochemistry and the Dental Research Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27514, U.S.A.
› Author Affiliations
Further Information

Publication History

Received 04 November 1976

Accepted 12 June 1977

Publication Date:
12 July 2018 (online)

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Summary

An improved method for the preparation of bovine α-thrombin is described. The procedure involves the activation of partially purified prothrombin with tissue thromboplastin followed by chromatography on Sulfopropyl-Sephadex C-50. The purified enzyme is homogeneous on polyacrylamide discontinuous gel electrophoresis and has a specific activity toward fibrinogen of 2,200–2,700 N.I.H. U/mg. Its stability on storage in liquid media is dependent on both ionic strenght and temperature. Increasing ionic strength and decreasing temperature result in optimal stability. The denaturation of α-thrombin by guanidine hydrochloride was found to be a partially reversible process with the renatured species possessing properties similar to “aged” thrombin. In addition, the catalytic properties of a-thrombin covalently attached to agarose gel beads were also examined. The activity of the immobilized enzyme toward fibrinogen was affected to a much greater extent than was the hydrolysis of low molecular weight, synthetic substrates.

 

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