PURIFICATION, CHARACTERIZATION, and AMINO-ACID-SEQUENCE of A SERINE PROTEINASE, PA-BJ, WITH PLATELET-AGGREGATING ACTIVITY FROM the VENOM of BOTHROPS-JARARACA

PURIFICATION, CHARACTERIZATION, and AMINO-ACID-SEQUENCE of A SERINE PROTEINASE, PA-BJ, WITH PLATELET-AGGREGATING ACTIVITY FROM the VENOM of BOTHROPS-JARARACA

Autor Serrano, SMT Google Scholar
Mentele, R. Google Scholar
Sampaio, CAM Google Scholar
Fink, E. Google Scholar
Instituição INST BUTANTAN
MAX PLANCK INST BIOCHEM
Universidade Federal de São Paulo (UNIFESP)
UNIV MUNICH
Resumo A platelet-aggregating enzyme, PA-BJ, was isolated from the venom of the snake Bothrops jararaca. PA-BJ in a concentration of 3.2 x 10(-8) M promoted 95% platelet aggregation in platelet-rich plasma. SDS-polyacrylamide gel electrophoresis under reducing conditions showed a single protein band with an M(r) of 30 000. PA-BJ catalyzed the hydrolysis of several p-nitroanilide peptide substrates containing Arg or Lys at the scissile bond; among these the most sensitive were the thrombin substrates D-Phe-Pip-Arg-pNA and Tos-Gly-Pro-Arg-pNA. Both the platelet-aggregating and amidolytic activities of PA-BJ were abolished by reaction with phenylmethanesulfonyl fluoride. Several benzamidine derivatives, which are competitive inhibitors of trypsin-like serine proteinases, also inhibited the amidolytic activity of PA-BJ. Among the compounds tested, the thrombin inhibitor NAPAP [((alpha)-[(2-naphthylsulfonyl)- glycyl]-4-amidinophenylalanine piperidide] showed the strongest inhibitor activity on PA-BJ. the complete amino acid sequence of PA-BJ, which, to the best of our knowledge, is the first of a platelet-aggregating enzyme from snake venom, was deduced from the N-terminal sequencing of overlapping fragments cleaved from the reduced and S-pyridylethylated protein by chemical and enzymatic methods. PA-BJ is composed of 232 amino acid residues and contains one N- and one O-glycosidically linked carbohydrate moiety at residues Asn(20) and Ser(23). Sequence comparison to other venom serine proteinases revealed significant homology, mainly in regions around the catalytic triad and conserved cysteine residues.
Idioma Inglês
Data de publicação 1995-05-30
Publicado em Biochemistry. Washington: Amer Chemical Soc, v. 34, n. 21, p. 7186-7193, 1995.
ISSN 0006-2960 (Sherpa/Romeo, fator de impacto)
Publicador Amer Chemical Soc
Extensão 7186-7193
Fonte http://dx.doi.org/10.1021/bi00021a033
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:A1995RA97600033
Endereço permanente http://repositorio.unifesp.br/handle/11600/25496

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