Myristate exchange on the Trypanosoma brucei variant surface glycoprotein.
The glycosyl-phosphatidylinositol (GPI) anchor of the Trypanosoma brucei variant surface glycoprotein (VSG) is unique in having exclusively myristate as its fatty acid component. We previously demonstrated that the myristate specificity is the result of two independent pathways. First, the newly synthesized free GPI, which is not myristoylated, undergoes fatty acid remodeling to replace both its fatty acids with myristate. Second, the myristoylated precursor, glycolipid A, undergoes a myristate exchange reaction, detected by the replacement of unlabeled myristate by [3H]myristate. Remodeling and exchange have different enzymatic properties and apparently occur in different subcellular compartments. We now demonstrate that the GPI anchor linked to VSG is the major substrate for myristate exchange. VSG can be efficiently labeled with [3H]myristate by exchange in the presence of cycloheximide, an inhibitor that prevents new VSG synthesis and thus anchor addition to protein. Not only is newly synthesized VSG subject to exchange, but mature VSG, possibly recycling from the cell surface, also undergoes myristate exchange.
Buxbaum
L U
LU
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Milne
K G
KG
Werbovetz
K A
KA
Englund
P T
PT
D000818:Animals; D001345:Autoradiography; D002474:Cell-Free System; D003513:Cycloheximide / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D007700:Kinetics; D008050:Lipid A / Q000378:metabolism; D019814:Myristic Acid; D009227:Myristic Acids / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D014316:Tritium; D014346:Trypanosoma brucei brucei / Q000187:drug effects / Q000378:metabolism*; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*
D017261:Glycosylphosphatidylinositols; D008050:Lipid A; D009227:Myristic Acids; D011500:Protein Synthesis Inhibitors; D014643:Variant Surface Glycoproteins, Trypanosoma; D019814:Myristic Acid; D014316:Tritium; D003513:Cycloheximide