title: Glycosylphosphatidylinositol anchors represent the major carbohydrate modification in proteins of intraerythrocytic stage Plasmodium falciparum.
abstract: The nature and extent of carbohydrate modification in intraerythrocytic stage Plasmodium falciparum proteins have been controversial. This study describes the characterization of the carbohydrates in intraerythrocytic P. falciparum proteins and provides an overall picture of the nature of carbohydrate modification in the parasite proteins. P. falciparum strains were metabolically labeled with radioactive sugar precursors and ethanolamine at different developmental stages. The individual parasite proteins separated on SDS-polyacrylamide gels and whole parasite cell lysates were analyzed for the carbohydrate moieties. The results established the following: 1) glycosylphosphatidylinositol (GPI) anchors represent the major carbohydrate modification in the intraerythrocytic stage P. falciparum proteins; 2) in contrast to previous reports, O-linked carbohydrates are either absent or present only at very low levels in the parasite; and 3) P. falciparum contains low levels of N-glycosylation capability. The amount of N-linked carbohydrates in whole parasite proteins is approximately 6% compared with the GPI anchors attached to proteins based on radioactive GlcN incorporated into the proteins. The glycan cores of multiple parasite protein GPI anchors are all similar, consisting of protein-ethanolamine-phosphate-(Manalpha1-2)6Manalpha1-2M analpha1-6Ma nalpha1- 4GlcN. The fourth Man residues distal to GlcN of the GPI anchor glycan cores contain unidentified substituents that are susceptible to conditions of nitrous acid deamination. This unusual structural feature may contribute to the reported pathogenic properties of the P. falciparum GPI anchors.
authors.affiliation: Department of Biochemistry and Molecular Biology, Georgetown University Medical Center, Washington, D. C. 20007, USA.
authors.forename: D C
authors.initials: DC
authors.lastname: Gowda
authors.forename: P
authors.initials: P
authors.lastname: Gupta
authors.forename: E A
authors.initials: EA
authors.lastname: Davidson
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D004912:Erythrocytes / Q000469:parasitology*; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols; D006595:Hexosamines / Q000032:analysis; D006801:Humans; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009608:Nitrous Acid / Q000737:chemistry; D010963:Plasmodium falciparum / Q000276:immunology / Q000378:metabolism*; D011134:Polysaccharides / Q000032:analysis; D011499:Protein Processing, Post-Translational; D015800:Protozoan Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
chemical_list: D017261:Glycosylphosphatidylinositols; D006595:Hexosamines; D008562:Membrane Glycoproteins; D011134:Polysaccharides; D015800:Protozoan Proteins; C025363:mannoproteins; D009608:Nitrous Acid