Characterization of carbohydrate binding proteins in Trypanosoma cruzi.
Trypanosoma cruzi is an obligatory intracellular protozoan parasite that causes Chagas' disease in humans and invades a great variety of mammalian cells. The nature of the ligand(s) and receptor components in both T. cruzi and target cells remains controversial, although it seems to involve an interaction with oligosaccharides. In an attempt to identify possible ligands on the parasite, we have searched for the presence of carbohydrate binding proteins (CBPs) in T. cruzi. By fluorescence-activated cell sorter analysis using a panel of fluoresceinated glyco- and neoglycopeptides with well characterized glycans, the presence of at least two different CBPs was identified on the surface of T. cruzi epimastigotes and trypomastigotes. The specificity of binding of the two CBPs seems to be mediated by galactose and mannose residues. The mannose- and galactose-mediated CBPs from epimastigotes and trypomastigotes were purified to homogeneity by affinity chromatography on immobilized thyroglobulin and identified as 60-70-kDa glycoproteins. Purified CBPs were able to specifically bind with high affinity to murine and human macrophages as well as other cell types susceptible to infection by T. cruzi but not to fat or neuronal cells. This binding was inhibited by the corresponding ligands. Moreover, the mannose-mediated CBP binding was completely abolished by alpha-mannosidase treatment of the cells. These results suggest a possible role for the CBPs in the recognition events between the parasite and target cells and/or in the interaction of the epimastigotes with the insect gut cells.
Bonay
P
P
Centro de Biologia Molecular, Universidad Autonoma de Madrid, Cantoblanco, Spain.
Fresno
M
M
D000818:Animals; D001212:Asialoglycoproteins / Q000378:metabolism; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D002846:Chromatography, Affinity; D003094:Collagen / Q000378:metabolism; D060748:Fetuins; D005434:Flow Cytometry; D006020:Glycopeptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D006493:Heparin / Q000378:metabolism; D006801:Humans; D007668:Kidney; D007700:Kinetics; D008969:Molecular Sequence Data; D011485:Protein Binding; D015800:Protozoan Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013954:Thyroglobulin / Q000031:analogs & derivatives / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000378:metabolism*; D014709:Vero Cells; D000509:alpha-Fetoproteins / Q000378:metabolism
D001212:Asialoglycoproteins; D060748:Fetuins; D006020:Glycopeptides; D015800:Protozoan Proteins; D000509:alpha-Fetoproteins; C017029:asialofetuin; C019149:asialothyroglobulin; D012710:Serum Albumin, Bovine; D006493:Heparin; D003094:Collagen; D013954:Thyroglobulin