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.
Laboratório de Ultra-estrutura Celular, IOC/FIOCRUZ, Av. Brasil 4365, Manguinhos, 21040-900 Rio de Janeiro, RJ, Brazil.
Francisco Odencio Rodrigues de
FO
Oliveira
Carlos Roberto
CR
Alves
Cláudia Magalhães
CM
Calvet
Leny
L
Toma
Rodrigo Ippolito
RI
Bouças
Helena Bociani
HB
Nader
Luzia Monteiro de
LM
Castro Côrtes
Marco Aurélio
MA
Krieger
Maria de Nazareth S L
Mde N
Meirelles
Mirian Claudia de
MC
Souza Pereira
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