Studies on O-glycans of Plasmodium-falciparum-infected human erythrocytes. Evidence for O-GlcNAc and O-GlcNAc-transferase in malaria parasites.
O-Glycosylation is the major form of protein glycosylation in human erythrocytes infected with the asexual intraerythrocytic stage of the malaria parasite. Plasmodium falciparum. This study compares aspects of O-glycosylation in P. falciparum-infected and uninfected erythrocytes. Non-labeled and metabolically glucosamine-labeled O-glycans were obtained from the protein fraction of infected or uninfected erythrocytes by beta elimination. Additional label was introduced by reduction with sodium borohydride, or by the attachment of radioactive Gal to peripheral GlcNAc using galactosyltransferase. 2-4-times more labeled O-glycans were obtained from infected erythrocytes compared to the same number of uninfected ones, consistent with additional biosynthesis by the parasite. Our analysis of these O-glycans showed no significant qualitative divergence between the O-glycans of the infected and those of the uninfected red cell. According to preliminary alditol analyses, the O-glycans of P. falciparum-infected red cells do not contain GalNAc at their reducing terminus. Moreover, GalNAc was not synthesized by P. falciparum from either Glc, Gal, GlcN or GalN. At least one O-glycan found in P. falciparum-infected erythrocytes contains GlcNAc at its reducing terminus. Gel-filtration results had suggested the presence of O-GlcNAc on proteins in the infected erythrocyte. Probing with a synthetic pentapeptide, we could show that P. falciparum expresses its own O-GlcNAc transferase during intraerythrocytic development. Using this peptide, the enzyme was characterized to some degree. The localization and function of O-GlcNAc in P. falciparum remains to be elucidated.
Dieckmann-Schuppert
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A
Zentrum für Hygiene und Medizinische Mikrobiologie, University of Marburg, Germany.
Bause
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E
Schwarz
R T
RT
D000117:Acetylglucosamine / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D002845:Chromatography; D004912:Erythrocytes / Q000378:metabolism* / Q000469:parasitology; D006801:Humans; D016778:Malaria, Falciparum / Q000097:blood* / Q000201:enzymology; D008969:Molecular Sequence Data; D017351:N-Acetylglucosaminyltransferases / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D010963:Plasmodium falciparum / Q000201:enzymology / Q000378:metabolism*; D011134:Polysaccharides / Q000097:blood* / Q000737:chemistry
D010455:Peptides; D011134:Polysaccharides; D017351:N-Acetylglucosaminyltransferases; D000117:Acetylglucosamine