Synthetic peptides and fluorogenic substrates related to the reactive site sequence of Kunitz-type inhibitors isolated from Bauhinia: Interaction with human plasma kallikrein

Synthetic peptides and fluorogenic substrates related to the reactive site sequence of Kunitz-type inhibitors isolated from Bauhinia: Interaction with human plasma kallikrein

Author Oliva, Maria Luiza Vilela Autor UNIFESP Google Scholar
Santomauro-Vaz, Eugenio Miguel Autor UNIFESP Google Scholar
Andrade, S. A. Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Pott, V. J. Google Scholar
Sampaio, Misako Uemura Autor UNIFESP Google Scholar
Sampaio, Claudio Augusto Machado Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
Abstract We have previously described Kunitz-type serine proteinase inhibitors purified from Bauhinia seeds. Human plasma kallikrein shows different susceptibility to those inhibitors. In this communication, we describe the interaction of human plasma kallikrein with fluorogenic and non-fluorogenic peptides based on the Bauhinia inhibitors' reactive site. The hydrolysis of the substrate based on the a. variegata inhibitor reactive site sequence, Abz-WISALPRSVFIQ-EDDnp (K-m 1.42 muM, k(cat) 0.06 s(-1), and k(cat)/K-m 4.23 x 10(4) M-1 s(-1)), is more favorable than that of Abz-VMIAALPRTMFIQ-EDDnp, related to the B. ungulata sequence (K-m 0.43 muM, k(cat) 0.00017 s(-1), and k(cat)/K-m 3.9 x 10(2) M-1 s(-1)). Human plasma kallikrein does not hydrolyze the substrates Abz-RPGLPVRFESPL-EDDnp and Abz-FESPLRINIIKE-EDDnp based on the a. bauhinioides inhibitor reactive site sequence, the most effective inhibitor of the enzyme. These peptides are competitive inhibitors with K-i values in the nM range. The synthetic peptide containing 19 amino acids based on the a. bauhinioides inhibitor reactive site (RPGLPVRFESPL) is poorly cleaved by kallikrein. The given substrates are highly specific for trypsin and chymotrypsin hydrolysis. Other serine proteinases such as factor Xa, factor XII, thrombin and plasmin do not hydrolyze a. bauhinioides inhibitor related substrates.
Keywords amino acid sequence
kallikrein
Kunitz inhibitor
peptide substrate
plasma kallikrein inhibitor
Language English
Date 2001-01-01
Published in Biological Chemistry. Berlin: Walter De Gruyter & Co, v. 382, n. 1, p. 109-113, 2001.
ISSN 1431-6730 (Sherpa/Romeo, impact factor)
Publisher Walter De Gruyter & Co
Extent 109-113
Origin https://doi.org/10.1515/BC.2001.016
Access rights Closed access
Type Article
Web of Science ID WOS:000167243100016
URI http://repositorio.unifesp.br/11600/42815

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