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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
Institution: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  d-Gal" target="_blank">d-Gal  d-galactose" target="_blank">d-galactose  d-Man" target="_blank">d-Man  d-mannose" target="_blank">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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