Human tissue kallikrein S-1 subsite recognition of non-natural basic amino acids

Human tissue kallikrein S-1 subsite recognition of non-natural basic amino acids

Autor Melo, R. L. Google Scholar
Pozzo, RCB Google Scholar
Pimenta, D. C. Google Scholar
Perissutti, E. Google Scholar
Caliendo, G. Google Scholar
Santagada, V Google Scholar
Juliano, L. Google Scholar
Juliano, M. A. Google Scholar
Instituição Universidade Federal de São Paulo (UNIFESP)
Univ Naples Federico II
Resumo We explored the unique substrate specificity of the primary S-1 subsite of human urinary kallikrein (hK1), which accepts both Phe and Arg, using internally quenched fluorescent peptides Abz-G-X-S-R-Q-EDDnp and Abz-G-F-S-P-F-X-S-S-R-P-B-EBBnp [Abz is o-aminobenzoic acid, EDDnp is N-(2,4-dinitrophenyl)ethylenediamine], which were based on the human kininogen sequence at the C-terminal region of bradykinin, Position X, which in natural sequence stands for Arg; received the following synthetic basic non-natural amino acids: 4-(aminomethyl)phenylalanine (Amf), 4-guanidine phenylalanine (Gnf), 4-(aminomethyl)-N-isopropylphenylala (Iaf), N-im-(dimethyl)histidine [H(2Me)], 3-pyridylalanine (Pya), 4-piperidinylalanine (Ppa), 4-(atninomethyl)cyclohexylalnnine (Ama), and 4-(aminocyclohexyl)alanine (Aca), Only Abz-F-Amf-S-R-Q-EDDnp and Abz-F-[H(2Me)]-S-R-B-EDDnp were efficiently hydrolyzed, and all others were resistant to hydrolysis. However, Abz-F-Ama-S-R-Q-EBDnp inhibited hK1 with a K-i of 50 nM with high specificity compared to human plasma kallikrein, thrombin, plasmin, and trypsin, the Abz-G-F-S-P-F-X-S-S-R-P-B-EDDnp series were more susceptible to hK1, although the peptides with Gnf, Ppa, and Ama were resistant to it. Unexpectedly, the peptides in which X is His, Lys, H(2Me), Amf, Iaf, Ppa, and Aca were cleaved at amino or at carboxyl sires of these amino acids, indicating that the S-1' subsite has significant preference for basic residues. Human plasma kallikrein did not hydrolyze any peptide of this series except the natural sequence where X is Arg. in conclusion, the Si subsite of hK1 accepts amino acids with combined basic anal aromatic side chain, although for the S-1-P-1 interaction the preference is for aliphatic and basic side chains.
Idioma Inglês
Data de publicação 2001-05-01
Publicado em Biochemistry. Washington: Amer Chemical Soc, v. 40, n. 17, p. 5226-5232, 2001.
ISSN 0006-2960 (Sherpa/Romeo, fator de impacto)
Publicador Amer Chemical Soc
Extensão 5226-5232
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000168435100014
Endereço permanente

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