Mycoplasma hyopneumoniae in vitro peptidase activities: Identification and cleavage of kallikrein-kinin system-like substrates

Mycoplasma hyopneumoniae in vitro peptidase activities: Identification and cleavage of kallikrein-kinin system-like substrates

Author Moitinho-Silva, Lucas Google Scholar
Kondo, Marcia Y. Autor UNIFESP Google Scholar
Oliveira, Lilian C. G. Autor UNIFESP Google Scholar
Okamoto, Debora N. Autor UNIFESP Google Scholar
Paes, Jessica A. Google Scholar
Machado, Mauricio F. M. Autor UNIFESP Google Scholar
Veronez, Camila L. Autor UNIFESP Google Scholar
Motta, Guacyara Autor UNIFESP Google Scholar
Andrade, Sheila S. Autor UNIFESP Google Scholar
Juliano, Maria A. Autor UNIFESP Google Scholar
Ferreira, Henrique B. Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Gouvea, Iuri E. Autor UNIFESP Google Scholar
Institution Univ Fed Rio Grande do Sul
Universidade Federal de São Paulo (UNIFESP)
Abstract Bacterial proteases are important for metabolic processes and pathogenesis in host organisms. the bacterial swine pathogen Mycoplasma hyopneumoniae has 15 putative protease-encoding genes annotated, but none of them have been functionally characterized. To identify and characterize peptidases that could be relevant for infection of swine hosts, we investigated the peptidase activity present in the pathogenic 7448 strain of M. hyopneumoniae. Combinatorial libraries of fluorescence resonance energy transfer peptides, specific inhibitors and pH profiling were used to screen and characterize endopeptidase, aminopeptidase and carboxypeptidase activities in cell lysates. One rnetalloendopeptidase, one serine endopeptidase, and one aminopeptidase were detected. the detected metalloendopeptidase activity, prominent at neutral and basic pH ranges, was due to a thimet oligopeptidase family member (M3 family), likely an oligoendopeptidase F (PepF), which cleaved the peptide Abz-GFSPFRQ-EDDnp at the F-S bond. A chymotrypsin-like serine endopeptidase activity, possibly a subtilisin-like serine protease, was prominent at higher pH levels, and was characterized by its preference for a Phe residue at the P-1 position of the substrate. the aminopeptidase P (APP) activity showed a similar profile to that of human membrane-bound APP. Genes coding for these three peptidases were identified and their transcription was confirmed in the 7448 strain. Furthermore, M. hyopneumoniae cell lysate peptidases showed effects on kallikrein-kinin system-like substrates, such as bradykinin-derived substrates and human high molecular weight kininogen. the M. hyopneumoniae peptidase activities, here characterized for the first time, may be important for bacterial survival strategies and thus represent possible targets for drug development against M. hyopneumoniae swine infections. (C) 2013 Elsevier B.V. All rights reserved.
Keywords Mycoplasma hyopneumoniae
Fluorescence resonance energy transfer peptides
Peptidase
Bradykinin
Kininogen
Language English
Sponsor Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Date 2013-05-03
Published in Veterinary Microbiology. Amsterdam: Elsevier B.V., v. 163, n. 3-4, p. 264-273, 2013.
ISSN 0378-1135 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 264-273
Origin http://dx.doi.org/10.1016/j.vetmic.2013.01.011
Access rights Closed access
Type Article
Web of Science ID WOS:000316926900007
URI http://repositorio.unifesp.br/handle/11600/36332

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