Autor |
Bersanetti, P. A.
![]() Andrade, MCC ![]() Casarini, D. E. ![]() Juliano, M. A. ![]() Nchinda, A. T. ![]() Sturrock, E. D. ![]() Juliano, L. ![]() Carmona, A. K. ![]() |
Instituição | Universidade Federal de São Paulo (UNIFESP) Univ Cape Town |
Resumo | Positional-scanning combinatorial libraries of fluorescence resonance energy transfer peptides were used for the analyses of the S-3 to S-1' subsites of the somatic angiotensin I-converting enzyme (ACE). Substrate specificity of ACE catalytic domains (C- and N-domains) was assessed in an effort to design selective substrates for the C-domain. Initially, we defined the S, specificity by preparing a library with the general structure Abz-GXXZXK(Dnp)-OH [Abz = o-aminobenzoic acid, K(Dnp) = N-epsilon-2,4-dinitrophenyllysine, and X is a random residue], where Z was successively occupied with one of the 19 natural amino acids with the exception of Cys. the peptides containing Arg and Leu in the P-1 position had higher C-domain selectivity. in the sublibraries Abz-GXXRZK(Dnp)-OH, Abz-GXZRXK(Dnp)-OH, and Abz-GZXRXK(Dnp)-OH, Arg was fixed at P-1 so we could define the C-domain selectivity of the S-1', S-2, and S-3 subsites. On the basis of the results from these libraries, we synthesized peptides Abz-GVIRFK(Dnp)-OH and Abz-GVILFK(Dnp)-OH which contain the most favorable residues for C-domain selectivity. Systematic reduction of the length of these two peptides resulted in Abz-LFK(Dnp)-OH, which demonstrated the highest selectivity for the recombinant ACE C-domain (k(cat)/K-m = 36.7 muM(-1) s(-1)) versus the N-domain (k(cat)/K-m = 0.51 muM(-1) s(-1)). the substrate binding of Abz-LFK(Dnp)-OH with testis ACE using a combination of conformational analysis and molecular docking was examined, and the results shed new light on the binding characteristics of the enzyme. |
Idioma | Inglês |
Data de publicação | 2004-12-21 |
Publicado em | Biochemistry. Washington: Amer Chemical Soc, v. 43, n. 50, p. 15729-15736, 2004. |
ISSN | 0006-2960 (Sherpa/Romeo, fator de impacto) |
Publicador | Amer Chemical Soc |
Extensão | 15729-15736 |
Fonte |
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Direito de acesso | Acesso restrito |
Tipo | Artigo |
Web of Science | WOS:000225776200008 |
Endereço permanente | http://repositorio.unifesp.br/handle/11600/28052 |
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