A novel fibrinogen-clotting enzyme, TL-BJ, from the venom of the snake Bothrops jararaca: Purification and characterization

A novel fibrinogen-clotting enzyme, TL-BJ, from the venom of the snake Bothrops jararaca: Purification and characterization

Author Serrano, Solange MT Google Scholar
Sampaio, Claudio AM Autor UNIFESP Google Scholar
Mentele, Reinhard Google Scholar
Camargo, Antonio CM Google Scholar
Fink, Edwin Google Scholar
Institution Inst Butantan
Universidade Federal de São Paulo (UNIFESP)
Univ Munich
Abstract Three chromatographically distinct forms of a novel fibrinogen-clotting 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 art: 16.8 NIH U/mg (TL-BJ 1), 16.7 NIH U/mg (TL-BJ 2) and 20.8 NLH 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-Arg-pNA 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 3 and TL-BJ 3 are identical while TL-BJ 1 has five substitutions.
Keywords snake venom
purification
serine peptidase
fibrinogen-clotting
Language English
Date 2000-03-01
Published in Thrombosis And Haemostasis. Stuttgart: F K Schattauer Verlag Gmbh, v. 83, n. 3, p. 438-444, 2000.
ISSN 0340-6245 (Sherpa/Romeo, impact factor)
Publisher F K Schattauer Verlag Gmbh
Extent 438-444
Access rights Closed access
Type Article
Web of Science ID WOS:000085876100016
URI http://repositorio.unifesp.br/11600/45659

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