Purification and characterization of a kinin-releasing and fibrinogen-clotting serine proteinase (KN-BJ) from the venom of Bothrops jararaca, and molecular closing and sequence analysis of its cDNA

Purification and characterization of a kinin-releasing and fibrinogen-clotting serine proteinase (KN-BJ) from the venom of Bothrops jararaca, and molecular closing and sequence analysis of its cDNA

Autor Serrano, SMT Google Scholar
Hagiwara, Y. Google Scholar
Murayama, N. Google Scholar
Higuchi, Z. Google Scholar
Mentele, R. Google Scholar
Sampaio, Claudio Augusto Machado Autor UNIFESP Google Scholar
Camargo, Antonio Carlos Martins de Autor UNIFESP Google Scholar
Fink, E. Google Scholar
Instituição Inst Butantan
Showa Univ
Univ Munich
Universidade Federal de São Paulo (UNIFESP)
Resumo Two forms of a proteinase, KN-BJ 1 and 2, were purified to homogeneity from the venom of Bothrops jararaca. in SDS/PAGE reduced KN-BJ 1 and 2 migrated as single bands with molecular masses of 38 kDa and 39 kDa. the two enzymes have similar N-terminal amino acid sequences and specific activities on synthetic chromogenic substrates, and both release bradykinin fi om bovine low-molecular-mass kininogen. KN-BJ 1 and KN-BJ 2 clot fibrinogen with specific activities of 245 NIH U/mg and 219 NIH U/mg, releasing only fibrinopeptide A. the amidolytic, kinin-releasing and coagulant activities are inhibited by phenylmethylsulfonyl fluoride, demonstrating that KN-BJ is a serine proteinase. Benzamidine derivatives, which are competitive inhibitors of trypsin-like proteinases. also inhibited the amidolytic activity of KN-BJ. A cDNA clone (HS104, 2.2 kb) has been isolated from a cDNA library of B. jararaca venom glands with an ORF of 771 bp. the deduced amino acid sequence contains segments that are identical to the sequences of the N-terminus and three tryptic peptides of KN-BJ 2. Therefore, the cDNA is believed to represent the gene of KN-BJ 2. the deduced amino acid sequence indicates that KN-BJ 2 is synthesized as a prezymogen of 257 amino acids with a putative signal peptide of 18 amino acids and an activating peptide of six amino acid residues. the sequence of 233 amino acids representing the mature enzyme exhibits high similarity to sequences of serine proteinases isolated from crotalid venoms.
Palavra-chave kinin
kallikrein
kininogenase
Bothrops jararaca
snake venom serine proteinase
Idioma Inglês
Data de publicação 1998-02-01
Publicado em European Journal of Biochemistry. New York: Springer Verlag, v. 251, n. 3, p. 845-853, 1998.
ISSN 0014-2956 (Sherpa/Romeo, fator de impacto)
Publicador Springer
Extensão 845-853
Fonte http://dx.doi.org/10.1046/j.1432-1327.1998.2510845.x
Direito de acesso Acesso aberto Open Access
Tipo Artigo
Web of Science WOS:000071855900036
Endereço permanente http://repositorio.unifesp.br/handle/11600/25857

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