Positional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides to define substrate specificity of carboxydipeptidases: assays with human cathepsin B

Positional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides to define substrate specificity of carboxydipeptidases: assays with human cathepsin B

Autor Cotrin, S. S. Google Scholar
Puzer, L. Google Scholar
Judice, WAD Google Scholar
Juliano, L. Google Scholar
Carmona, A. K. Google Scholar
Juliano, M. A. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Resumo We have developed positional scanning synthetic combinatorial libraries to define the substrate specificity of carboxydipeptidases. the library Abz-GXXZXK(Dnp)-OH, where Abz is ortho-aminobenzoic acid, K(Dnp) is N-c-2,4-dinitrophenyl-lysine with free carboxyl group, the Z position was successively occupied with 1 of 19 amino acids (eysteine was omitted), and X represents randomly incorporated residues, was assayed initially with human cathepsin B, and arginine was defined as one of the best residues at the P, position. To examine the selectivity of S-1(1) S-2, and S-3 subsites, the sublibraries Abz-GXXRZK(Dnp)-OH, AbzGXZRXK(Dnp)-OH, and Abz-GZXRXK(Dnp)-OH were then synthesized. the peptide Abz-GIVRAK(Dnp)-OH, which contains the most favorable residues in the P-3-P-1, positions identified by screening of the libraries with cathepsin B, was hydrolyzed by this enzyme with k(cat)/K-m = 7288 mM(-1) s(-1). This peptide is the most efficient substrate described for cathepsin B to this point, and it is highly selective for the enzyme among the lysosomal cysteine proteases. (C) 2004 Elsevier Inc. All rights reserved.
Palavra-chave cysteine peptidase
lysosome
amino acids
exopeptidase
Idioma Inglês
Data de publicação 2004-12-15
Publicado em Analytical Biochemistry. San Diego: Academic Press Inc Elsevier Science, v. 335, n. 2, p. 244-252, 2004.
ISSN 0003-2697 (Sherpa/Romeo, fator de impacto)
Publicador Elsevier B.V.
Extensão 244-252
Fonte http://dx.doi.org/10.1016/j.ab.2004.09.012
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000225502800009
Endereço permanente http://repositorio.unifesp.br/handle/11600/28050

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