Conserved cystatin segments as models for designing specific substrates and inhibitors of cysteine proteinases

Conserved cystatin segments as models for designing specific substrates and inhibitors of cysteine proteinases

Autor Lalmanach, G. Google Scholar
Serveau, C. Google Scholar
BrillardBourdet, M. Google Scholar
Chagas, JR Google Scholar
Mayer, R. Google Scholar
Juliano, L. Google Scholar
Gauthier, F. Google Scholar
Instituição UNIV TOURS
Universidade Federal de São Paulo (UNIFESP)
Resumo Peptide segments derived from consensus sequences of the inhibitory site of cystatins, the natural inhibitors of cysteine proteinases, were used to develop new substrates and inhibitors of papain and rat liver cathepsins B, H, and L. Papain hydrolyzed Abz-QVVAGA-EDDnp and Abz-LVGGA-EDDnp at about the same rate, with specificity constants in the 10(7) M(-1) sec(-1) range; cathepsin L also hydrolyzes both substrates with specificity constants in the 10(5) M(-1) sec(-1) range due to lower k(cat) values, with the K-m's being identical to those with papain. Only Abz-LVGGA-EDDnp was rapidly hydrolyzed by cathepsin B, and to a lesser extent by cathepsin H. Peptide substrates that alternate these two building blocks (LVGGQVVAGAPWK and QVVAGALVGGAPWK) discriminate the activities of cathepsins B and L and papain. Cathepsin L was highly selective for cleavage at the G-G bond of the LVGG fragment in both peptides. Papain and cathepsin B cleaved either the LVGG fragment or the QVVAG fragment, depending an their position within the peptide. While papain was more specific for the segment located C-terminally, cathepsin B was specific for that in N-terminal position. Peptidyl diazomethylketone inhibitors based an these two sequences also reacted differently with papain and cathepsins. GlcA-QVVA-CHN, was a potent inhibitor of papain and reacted with papain 60 times more rapidly (k(+0) = 1,100,000 M(-1) sec(-1)) than with cathepsin L, and 220 times more rapidly than with cathepsin B. Cathepsins B and L were preferentially inhibited by Z-RLVG-CHN2. Thus cystatin-derived peptides provide a valuable framework for designing sensitive, selective substrates and inhibitors of cysteine proteinases.
Palavra-chave cysteine proteinase
peptidyl fluorogenic substrate
Idioma Inglês
Data de publicação 1995-11-01
Publicado em Journal of Protein Chemistry. New York: Plenum Publ Corp, v. 14, n. 8, p. 645-653, 1995.
ISSN 0277-8033 (Sherpa/Romeo, fator de impacto)
Publicador Plenum Publ Corp
Extensão 645-653
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:A1995TM17500002
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