Characterization of a glycosyl-phosphatidylinositol-anchored membrane protein from Trypanosoma cruzi.
Four monoclonal antibodies (MAbs) specific for Trypanosoma cruzi were obtained. Flow cytometry analysis showed that these four MAbs stained the membranes of the three main morphological forms of T. cruzi: amastigotes, trypomastigotes, and epimastigotes. The four MAbs seemed to recognize the same 50- to 55-kDa antigen that was revealed by immunoblotting. Competition experiments revealed that they defined at least two different epitopes on the molecule. The antigen was detected on the external surface of the membrane by immunoelectron microscopy. Several experiments indicated that the 50- to 55-kDa antigen recognized by these four MAbs was a glycosyl-phosphatidylinositol-anchored membrane protein. (i) The antigen could be removed from the cell surface by treatment with proteases, NaOH, HNO2, and phosphatidylinositol-specific phospholipase C (PI-PLC). (ii) The phase distribution of the antigen in Triton X-114 solutions changed drastically upon treatment with PI-PLC. The antigen was found mainly in the detergent phase in nontreated samples and in the aqueous phase in PI-PLC-digested samples. (iii) A cross-reacting determinant that was found in other glycosyl-phosphatidylinositol-anchored membrane proteins appeared after PI-PLC treatment.
Hernández-Munaín
C
C
Centro de Biología Molecular Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Spain.
Fernández
M A
MA
Alcina
A
A
Fresno
M
M
D000818:Animals; D000911:Antibodies, Monoclonal; D006017:Glycolipids / Q000032:analysis* / Q000276:immunology; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D010716:Phosphatidylinositols / Q000032:analysis* / Q000276:immunology; D014349:Trypanosoma cruzi / Q000032:analysis*
D000911:Antibodies, Monoclonal; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D008565:Membrane Proteins; D010716:Phosphatidylinositols