Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/10681
Title: Isolation and molecular characterization of a major hemolymph serpin from the triatomine, Panstrongylus megistus
Authors: Moreira, Carlos J. C.
Waniek, Peter J.
Valente, Richard H.
Carvalho, Paulo C.
Perales, Jonas
Feder, Denise
Geraldo, Reinaldo B.
Castro, Helena C.
Azambuja, Patricia
Ratclife, Norman A.
Mello, Cícero B.
Affilliation: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Doenças Parasitárias. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxonologia. Rio de Janeiro, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Proteômica e Engenharia de Proteínas. Curitiba, PR, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Toxonologia. Rio de Janeiro, RJ, Brasil.
Universidade Federal Fluminense. Laboratório de Biologia de Insetos. Niterói, Brasil.
Universidade Federal Fluminense.IB. GCM. Laboratório LABIEMol. Niterói, RJ, Brasil.
Universidade Federal Fluminense.IB. GCM. Laboratório LABIEMol. Niterói, RJ, Brasil.
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Bioquímica e Fisiologia de Insetos. Rio de Janeiro, RJ, Brasil.
Universidade Federal Fluminense. Laboratório de Biologia de Insetos. Niterói, Brasil / Swansea University. College of Science. Department od Biosciences. Wales, UK.
Universidade Federal Fluminense. Laboratório de Biologia de Insetos. Niterói, Brasil.
Abstract: Background: Chagas disease kills 2.5 thousand people per year of 15 million persons infected in Latin America. The disease is caused by the protozoan, Trypanosome cruzi, and vectored by triatomine insects, including Panstrongylus megistus, an important vector in Brazil. Medicines treating Chagas disease have unpleasant side effects and may be ineffective, therefore, alternative control techniques are required. Knowledge of the T. cruzi interactions with the triatomine host needs extending and new targets/strategies for control identified. Serine and cysteine peptidases play vital roles in protozoan life cycles including invasion and entry of T. cruzi into host cells. Peptidase inhibitors are, therefore, promising targets for disease control. Methods: SDS PAGE and chromatograpy detected and isolated a P. megistus serpin which was peptide sequenced by mass spectrometry. A full amino acid sequence was obtained from the cDNA and compared with other insect serpins. Reverse transcription PCR analysis measured serpin transcripts of P. megistus tissues with and without T. cruzi infection. Serpin homology modeling used the Swiss Model and Swiss-PDB viewer programmes. Results: The P. megistus serpin (PMSRP1) has a ca. 40 kDa molecular mass with 404 amino acid residues. A reactive site loop contains a highly conserved hinge region but, based on sequence alignment, the normal cleavage site for serine proteases at P1-P1′ was translocated to the putative position P4′-P5′. A small peptide obtained corresponded to the C-terminal 40 amino acid region. The secondary structure of PMSRP1 indicated nine α-helices and three β-sheets, similar to other serpins. PMSRP1 transcripts occurred in all tested tissues but were highest in the fat body and hemocytes. Levels of mRNA encoding PMSRP1 were significantly modulated in the hemocytes and stomach by T. cruzi infection indicating a role for PMSRP1 in the parasite interactions with P. megistus. Conclusions: For the first time, a constitutively expressed serpin has been characterized from the hemolymph of a triatomine. This opens up new research avenues into the roles of serine peptidases in the T. cruzi/P. megistus association. Initial experiments indicate a role for PMSRP1 in T. cruzi interactions with P. megistus and will lead to further functional studies of this molecule.
Keywords: Panstrongylus megistus
Serpin
Hemolymph
Serine protease
Cleavage sites
T. cruzi serpin modulation
DeCS: Trypanosome cruzi
Doença de Chagas
Serpinas
Issue Date: 2014
Publisher: BioMed Central
Citation: MOREIRA, Carlos J. C. et al. Isolation and molecular characterization of a major hemolymph serpin from the triatomine, Panstrongylus megistus. Parasite & Vectors, v. 23, n. 7, p. 1-16, 2014.
DOI: 10.1186/1756-3305-7-23
ISSN: 1756-3305
Copyright: open access
Appears in Collections:PR - ICC - Artigos de Periódicos
IOC - Artigos de Periódicos

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