Specificity of cathepsin B to fluorescent substrates containing benzyl side-chain-substituted amino acids at P1 subsite

Specificity of cathepsin B to fluorescent substrates containing benzyl side-chain-substituted amino acids at P1 subsite

Autor Del Nery, E. Google Scholar
Alves, L. C. Google Scholar
Melo, R. L. Google Scholar
Cesari, MHS Google Scholar
Juliano, L. Google Scholar
Juliano, M. A. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Resumo We have determined the kinetic parameters for the hydrolysis by cathepsin B of peptidyl-coumarin amide and intramolecularly quenched fluorogenic peptides with the general structures epsilon NH2-Cap-Leu-X-MCA and Abz-Lys-Leu-X-Phe-Ser-Lys-Gln-EDDnp, respectively. Abz (ortho-aminobenzoic acid) and EDDnp (2,4-dinitrophenyl-ethylenediamine) are the fluorescent donor-acceptor pair, and X was Cys(SBzl), Ser(OBzl), and Thr(OBzl) containing benzyl group (Bzl) at the functional side chain of Cys, Ser, and Thr. the peptidyl-coumarin-containing Cys(SBzl), Ser(OBzl), and Thr(OBzl) have higher affinity cathepsin B, supporting the interpretation of the crystal structure of rat cathepsin B complexed with the inhibitor Z-Arg-Ser(OBzl)-CH2Cl that the benzyl group attached to Ser hydroxyl side chain occupies the enzyme S'(1) subsite [Jia et al. (1995), J. Biol. Chem. 270, 5527]. A similar effect of benzyl group was also detected in the internally quenched peptides. Finally, the benzyl group in substrates containing Cys(SBzl) amino acid at P-1 seems to compensate the absence of adequate S-2-P-2 interaction in the hydrolysis of the peptides having Pro or Ala at P-2 position.
Palavra-chave thiolprotease
cathepsins B
papain
fluorogenic substrate
fluorescent peptides
proteinases
Idioma Inglês
Data de publicação 2000-01-01
Publicado em Journal of Protein Chemistry. New York: Kluwer Academic/plenum Publ, v. 19, n. 1, p. 33-38, 2000.
ISSN 0277-8033 (Sherpa/Romeo, fator de impacto)
Publicador Kluwer Academic/plenum Publ
Extensão 33-38
Fonte http://dx.doi.org/10.1023/A:1007090708945
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000087777700004
Endereço permanente http://repositorio.unifesp.br/handle/11600/26224

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