Glycolipid precursors for the membrane anchor of Trypanosoma brucei variant surface glycoproteins. I. Can structure of the phosphatidylinositol-specific phospholipase C sensitive and resistant glycolipids.
A number of eukaryotic surface glycoproteins, including the variant surface glycoproteins of Trypanosoma brucei, are synthesized with a carboxyl-terminal hydrophobic peptide extension that is cleaved and replaced by a complex glycosyl-phosphatidylinositol (GPI) membrane anchor within 1-5 min of the completion of polypeptide synthesis. The rapidity of this carboxyl-terminal modification suggests the existence of a prefabricated precursor glycolipid that can be transferred en bloc to the polypeptide. We have reported the purification and partial characterization of a candidate precursor glycolipid (P2) and of a compositionally similar glycolipid (P3) from T. brucei (Menon, A. K., Mayor, S., Ferguson, M. A. J., Duszenko, M., and Cross, G. A. M. (1988) J. Biol. Chem. 263, 1970-1977). The primary structure of the glycan portions of P2 and P3 have now been analyzed by a combination of selective chemical fragmentation and enzymatic glycan sequencing at the subnanomolar level. The glycans were generated by deamination, NaB3H4 reduction, and dephosphorylation of glycolipids purified from different trypanosome variants. Glycan fragments derived from biosynthetically labeled glycolipids were also analyzed. The cumulative data strongly suggest that P2 and P3 contain ethanolamine-phosphate-Man alpha 1-2Man alpha 1-6Man alpha 1-GlcN linked glycosidically to an inositol residue, as do all the GPI anchors that have been structurally characterized. The structural similarities suggest that GPI membrane anchors are derived from common precursor glycolipids that become variably modified during or after addition to newly synthesized proteins.
Mayor
S
S
Rockefeller University, New York, New York 10021.
Menon
A K
AK
Cross
G A
GA
Ferguson
M A
MA
Dwek
R A
RA
Rademacher
T W
TW
D000818:Animals; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D006017:Glycolipids / Q000302:isolation & purification / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols; D007294:Inositol / Q000378:metabolism; D007700:Kinetics; D008563:Membrane Lipids / Q000378:metabolism*; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000302:isolation & purification / Q000378:metabolism*; D011134:Polysaccharides / Q000302:isolation & purification; D014346:Trypanosoma brucei brucei / Q000276:immunology / Q000378:metabolism*; D010738:Type C Phospholipases / Q000378:metabolism*; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000096:biosynthesis*
D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D008563:Membrane Lipids; D010716:Phosphatidylinositols; D011134:Polysaccharides; D014643:Variant Surface Glycoproteins, Trypanosoma; D007294:Inositol; D010738:Type C Phospholipases