A Kazal-type inhibitor is modulated by Trypanosoma cruzi to control microbiota inside the anterior midgut of Rhodnius prolixus

Biochimie
Tatiane S SoaresAparecida S Tanaka

Abstract

The triatomine insect, Rhodnius prolixus, is a vector of Trypanosoma cruzi, a protozoan parasite that causes Chagas disease. The parasite must overcome immune response and microbiota to develop inside the midgut of triatomines. In this study, we expressed, purified and characterized a Kazal-type inhibitor from the midgut of R. prolixus, named RpTI, which may be involved in microbiota - T. cruzi interactions. The qPCR showed that the RpTI transcript was primarily expressed in tissues from the intestinal tract and that it was upregulated in the anterior midgut after T. cruzi infection. A 315-bp cDNA fragment encoding the mature protein was cloned into the pPIC9 vector and expressed in Pichia pastoris system. Recombinant RpTI (rRpTI) was purified on a trypsin-Sepharose column and had a molecular mass of 11.5 kDa as determined by SDS-PAGE analysis. This protein inhibited trypsin (Ki = 0.42 nM), whereas serine proteases from the coagulation cascade were not inhibited. Moreover, trypanocidal assays revealed that rRpTI did not interfere in the viability of T. cruzi trypomastigotes. The RpTI transcript was also knocked down by RNA interference prior to infection of R. prolixus with T. cruzi. The amount of T. cruzi in the anterior midgu...Continue Reading

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Jan 29, 2016·Parasitology·Roberta Carvalho FerreiraAlessandra Aparecida Guarneri
Jul 13, 2016·Journal of Insect Physiology·Alessandra Aparecida Guarneri, Marcelo Gustavo Lorenzo
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Sep 14, 2018·PLoS Neglected Tropical Diseases·Michele Souza LimaGeorgia Correa Atella
Sep 7, 2019·Microbial Pathogenesis·Isabella Márcia Soares Nogueira TeotônioMariana Hecht
Jun 8, 2021·Research and Reports in Tropical Medicine·Günter A Schaub

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