P-I class metalloproteinase from Bothrops moojeni venom is a post-proline cleaving peptidase with kininogenase activity: Insights into substrate selectivity and kinetic behavior

P-I class metalloproteinase from Bothrops moojeni venom is a post-proline cleaving peptidase with kininogenase activity: Insights into substrate selectivity and kinetic behavior

Author Okamoto, Debora Noma Autor UNIFESP Google Scholar
Kondo, Marcia Yuri Autor UNIFESP Google Scholar
Oliveira, Lilian Caroline Gonçalves de Autor UNIFESP Google Scholar
Honorato, Rodrigo Vargas Google Scholar
Zanphorlin, Letícia Maria Google Scholar
Coronado, Monika Aparecida Google Scholar
Araujo, Mariana Silva Autor UNIFESP Google Scholar
Motta, Guacyara da Autor UNIFESP Google Scholar
Veronez, Camila Lopes Autor UNIFESP Google Scholar
Andrade, Sheila Siqueira Autor UNIFESP Google Scholar
Oliveira, Paulo Sergio Lopes de Google Scholar
Arni, Raghuvir Krishnaswamy Google Scholar
Cintra, Adélia Cristina Oliveira de Google Scholar
Sampaio, Suely Vilela Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Murakami, Mario Tyago Google Scholar
Gouvea, Iuri Estrada Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Ctr Nacl Pesquisas Energia & Mat
Universidade Estadual de Campinas (UNICAMP)
Universidade de São Paulo (USP)
Abstract Snake venom metalloproteinases (SVMPs) belonging to P-I class are able to hydrolyze extracellular matrix proteins and coagulation factors triggering local and systemic reactions by multiple molecular mechanisms that are not fully understood. BmooMP alpha-I, a P-I class SMVP from Bothrops moojeni venom, was active upon neuro- and vaso-active peptides including angiotensin I, bradykinin, neurotensin, oxytocin and substance P. Interestingly, BmooMPa-I showed a strong bias towards hydrolysis after proline residues, which is unusual for most of characterized peptidases. Moreover, the enzyme showed kininogenase activity similar to that observed in plasma and cells by kallikrein. FRET peptide assays indicated a relative promiscuity at its S-2-S '(2) subsites, with proline determining the scissile bond. This unusual post-proline cleaving activity was confirmed by the efficient hydrolysis of the synthetic combinatorial library MCA-GXXPXXQ-EDDnp, described as resistant for canonical peptidases, only after Pro residues. Structural analysis of the tripeptide LPL complexed with BmooMP alpha-I, generated by molecular dynamics simulations, assisted in defining the subsites and provided the structural basis for subsite preferences such as the restriction of basic residues at the S-2 subsite due to repulsive electrostatic effects and the steric impediment for large aliphatic or aromatic side chains at the Si subsite. These new functional and structural findings provided a further understanding of the molecular mechanisms governing the physiological effects of this important class of enzymes in envenomation process. (c) 2014 Elsevier B.V. All rights reserved.
Keywords Snake venom metalloproteinase
Kininogenase activity
FRET peptides
Substrate specificity
Molecular dynamics simulations
Language English
Sponsor Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Grant number FAPESP: 2012/50191-4
Date 2014-03-01
Published in Biochimica Et Biophysica Acta-proteins and Proteomics. Amsterdam: Elsevier B.V., v. 1844, n. 3, p. 545-552, 2014.
ISSN 1570-9639 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 545-552
Origin http://dx.doi.org/10.1016/j.bbapap.2013.12.014
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000333491100008
URI http://repositorio.unifesp.br/handle/11600/37451

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