Trypanosoma cruzi heparin-binding proteins and the nature of the host cell heparan sulfate-binding domain.
Trypanosoma cruzi invasion is mediated by receptor-ligand recognition between the surfaces of both parasite and target cell. We have previously demonstrated the role of heparan sulfate proteoglycan in the attachment and invasion of T. cruzi in cardiomyocytes. Herein, we have isolated the T. cruzi heparin-binding proteins (HBP-Tc) and investigated the nature of cardiomyocyte heparan sulfate (HS)-binding site to the parasite surface ligand. Two major heparin-binding proteins with molecular masses of 65.8 and 59 kDa were observed in total extract of amastigote and trypomastigote forms of T. cruzi. Hydrophobic [S(35)]methionine labeled proteins eluted from heparin-sepharose affinity chromatography also revealed both proteins in trypomastigotes but only the 59 kDa is strongly recognized by biotin-conjugated glycosaminoglycans. Competition assays were performed to analyze the role of sulfated proteoglycans, including heparin, keratan sulfate and both acetylated and highly sulfated domains of heparan sulfate, in the recognition and invasion process of T. cruzi. Significant inhibitions of 84% and 35% in the percentage of infection were revealed after treatment of the parasites with heparin and the N-acetylated/ N-sulfated heparan sulfate domain, respectively, suggesting the important role of the glycuronic acid and NS glucosamine domain of the HS chain in the recognition of the HBP-Tc during the T. cruzi-cardiomyocyte interaction.
Oliveira
Francisco Odencio Rodrigues de
FO
Laboratório de Ultra-estrutura Celular, IOC/FIOCRUZ, Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil.
Alves
Carlos Roberto
CR
Calvet
Cláudia Magalhães
CM
Toma
Leny
L
Bouças
Rodrigo Ippolito
RI
Nader
Helena Bociani
HB
Castro Côrtes
Luzia Monteiro de
LM
Krieger
Marco Aurélio
MA
Meirelles
Maria de Nazareth S L
Mde N
Souza Pereira
Mirian Claudia de
MC
D000818:Animals; D002478:Cells, Cultured; D002704:Chlorates / Q000494:pharmacology; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000469:parasitology; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D014349:Trypanosoma cruzi / Q000187:drug effects / Q000378:metabolism*; D014709:Vero Cells
D002704:Chlorates; D019812:Heparan Sulfate Proteoglycans; D015800:Protozoan Proteins; D006493:Heparin; C032706:sodium chlorate