Characterization of a phospholipase C-resistant inositol containing glycolipid from Trypanosoma cruzi.
Since glycosylphosphatidylinositol is the most common form of attachment of proteins to membranes in T. cruzi, and that this parasite depends on surface-mediated interactions for survival within the vector and mammalian host, it is probable that a drug which interfers with the metabolism of glycosylphosphatidylinositol (GPI) could be successfully employed in chemotherapy. Over the last few years several groups have been characterizing this mode of attachment in T. cruzi and more recently we have been concentrating our efforts on the identification of candidate precursors for protein anchors in metacyclic trypomastigotes. Previously detected GPI heterogeneity regarding solubilization of a major stage-specific antigen (1G7-Ag) by phospholipase C led us to investigate whether biosynthetic precursors with similar properties could also be identified. Two glycolipid species whose migration properties resemble glycolipids A and C of T. brucei were amenable to biosynthetic radiolabelling with palmitic acid, inositol, ethanolamine, glucosamine and mannose. Following purification, these species were submitted to classical GPI diagnostic treatments. In both cases digestion with GPI-specific phospholipase D (GPIPLD) produced phospatidic acid and treatment with either mild base or phospholipase A2 (PLA2) produced free fatty acid, indicating an acylation at least at position 2 of the glycerol. The glycolipid A-like species proved to be susceptible to solubilization by PIPLC of B. thuringiensis and by GPIPLC of T. brucei and the glycolipid C-like material proved to be fully resistant to both lipases.(ABSTRACT TRUNCATED AT 250 WORDS)
Heise
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Departamento de Microbiologia, Imunologia e Parasitologia, Escola Paulista de Medicina, São Paulo, Brasil.
Raper
J
J
Cardoso-de-Almeida
M L
ML
D000818:Animals; D002855:Chromatography, Thin Layer; D005227:Fatty Acids / Q000737:chemistry* / Q000378:metabolism; D006017:Glycolipids / Q000737:chemistry* / Q000378:metabolism; D017261:Glycosylphosphatidylinositols / Q000737:chemistry* / Q000378:metabolism; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry* / Q000378:metabolism; D014349:Trypanosoma cruzi / Q000737:chemistry* / Q000378:metabolism; D010738:Type C Phospholipases / Q000737:chemistry* / Q000378:metabolism
D005227:Fatty Acids; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D011498:Protein Precursors; D015800:Protozoan Proteins; D010738:Type C Phospholipases