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Thromb Haemost 2000; 83(03): 438-444
DOI: 10.1055/s-0037-1613834
Review Article
Schattauer GmbH

A Novel Fibrinogen-clotting Enzyme, TL-BJ, from the Venom of the Snake Bothrops jararaca: Purification and Characterization

Solange M. T. Serrano
1   From the Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo
,
Claudio A. M. Sampaio
2   From the Departamento de Bioquímica, EPM-UNIFESP, São Paulo, Brazil
,
Reinhard Mentele
3   From the Abteilung für Klinische Chemie und Klinische Biochemie, Chirurgische Klinik, Klinikum Innenstadt, Ludwig-Maximilians-Universität, München, Germany
,
Antonio C. M. Camargo
1   From the Laboratório de Bioquímica e Biofísica, Instituto Butantan, São Paulo
,
Edwin Fink
3   From the Abteilung für Klinische Chemie und Klinische Biochemie, Chirurgische Klinik, Klinikum Innenstadt, Ludwig-Maximilians-Universität, München, Germany
› Author Affiliations
Further Information

Publication History

Received 05 April 1999

Accepted after resubmission 19 November 1999

Publication Date:
14 December 2017 (online)

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Summary

Three chromatographically distinct forms of a novel fibrinogenclotting serine endopeptidase, TL-BJ 1, 2 and 3, were purified from the venom of Bothrops jararaca by a combination of ammonium sulfate precipitation and chromatographic steps. The three forms of TL-BJ have similar amidolytic and plasma coagulating activities. TL-BJ 1, TL-BJ 2 and TL-BJ 3 cause the specific clotting of fibrinogen with release of fibrinopeptide A, the specific activities are 16.8 NIH U/mg (TL-BJ 1), 16.7 NIH U/mg (TL-BJ 2) and 20.8 NIH U/mg (TL-BJ 3). The most sensitive chromogenic substrates for measuring the amidolytic activity of TL-BJ 3 were D-Pro-Phe-Arg-pNA, D-Phe-pipecolyl-ArgpNA and Z-D-Arg-Gly-Arg-pNA. The amidolytic and coagulant activities of TL-BJ were inhibited by phenylmethylsulfonyl fluoride but not by hirudin. Benzamidine derivatives, which are competitive inhibitors of trypsin-like serine endopeptidases, also inhibited the amidolytic activity of TL-BJ. In SDS/PAGE the main bands of TL-BJ 1, 2 and 3 showed molecular masses of 30 kDa, 31 kDa and 32 kDa. Upon incubation with N-glycosidase F only TL-BJ 3 remained unchanged, whereas TL-BJ 1 and TL-BJ 2 showed products with molecular masses around 23 kDa. Thus, TL-BJ 3 does not seem to be N-glycosylated. The N-terminal amino acid sequences of TL-BJ 2 and TL-BJ 3 are identical while TL-BJ 1 has five substitutions.

Keywords

Snake venom - purification - serine peptidase - fibrinogen-clotting
 

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