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Trypanosoma cruzi: attachment to perimicrovillar membrane glycoproteins of Rhodnius prolixus
Authors:Alves C R  Albuquerque-Cunha J M  Mello C B  Garcia E S  Nogueira N F  Bourguingnon S C  de Souza W  Azambuja P  Gonzalez M S
Affiliation:Laboratório de Biologia Molecular e Doen?as Endêmicas, Departamento de Bioquímica e Biologia Molecular, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil.
Abstract:Studies were carried out to identify proteins involved in the interface of Trypanosoma cruzi with the perimicrovillar membranes (PMM) of Rhodnius prolixus. Video microscopy experiments demonstrated high level of adhesion of T. cruzi Dm 28c epimastigotes to the surface of posterior midgut cells of non-treated R. prolixus. The parasites however were unable to attach to gut cells obtained from decapitated or azadirachtin-treated insects. The influence of carbohydrates on the adhesion to insect midgut was confirmed by inhibition of parasite attachment after midgut incubation with N-acetylgalactosamine, N-acetylmannosamine, N-acetylglucosamine, D-galactose, D-mannose or sialic acid. We observed that hydrophobic proteins in the surface of epimastigotes bind to polypeptides with 47.7, 45.5, 44, 43, 40.5, 36, 31 and 13kDa from R. prolixus PMM and that pre-incubation of lectins specifically inhibited binding to 31, 40.5, 44 and 45.5kDa proteins. We suggest that glycoproteins from PMM and hydrophobic proteins from epimastigotes are important for the adhesion of the parasite to the posterior midgut cells of the vector.
Keywords:Chagas disease   Rhodnius prolixus (Hemiptera:Reduviidae)   Triatomine vector   Trypanosoma cruzi (Kinetoplastida: Trypanossomatidae)   Peritrophic membranes   Hydrophobic proteins   Lectins   PMM, perimicrovillar membranes   GalNAc, N-acetylgalactosamine   ManNAc, N-acetylmannosamine   GlcNAc, N-acetylglucosamine     smallcaps"  >d-Gal,   smallcaps"  >d-galactose     smallcaps"  >d-Man,   smallcaps"  >d-mannose   NANA, sialic acid   WGA, wheat germ agglutinin   PNA, peanut agglutinin   Con A, concanavalin A   PTTH, protoracicotropic hormone   HCP, head critical period   CBPs, carbohydrate-binding proteins   PBS, phosphate-buffered saline   FCS, heat-inactivated fetal calf serum 20%   BHI, brain heart infusion culture medium   TX-114, Triton X-114   Rf, relative mobility
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