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PMID: 7621823 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The role of inositol acylation and inositol deacylation in GPI biosynthesis in Trypanosoma brucei.

The EMBO journal ·Vol. 14 ·No. 13 ·1995-07-03 ·Pages 3080-93

Güther ML, Ferguson MA

Abstract

The compound diisopropylfluorophosphate (DFP) selectively inhibits an inositol deacylase activity in living trypanosomes that, together with the previously described phenylmethylsulfonyl fluoride (PMSF)-sensitive inositol acyltransferase, maintains a dynamic equilibrium between the glycosylphosphatidylinositol (GPI) anchor precursor, glycolipid A [NH2(CH2)2PO4-6Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcN alpha 1-6myo-inositol-1-PO4-sn-1,2-dimyristoylglycerol], and its inositol acylated form, glycolipid C. Experiments using DFP in living trypanosomes and a trypanosome cell-free system suggest that earlier GPI intermediates are also in equilibrium between their inositol acylated and nonacylated forms. However, unlike mammalian and yeast cells, bloodstream form trypanosomes do not appear to produce an inositol acylated form of glucosaminylphosphatidylinositol (GlcN-PI). A specific function of inositol acylation in trypanosomes may be to enhance the efficiency of ethanolamine phosphate addition to the Man3GlcN-(acyl)PI intermediate. Inositol deacylation appears to be a prerequisite for fatty acid remodelling of GPI intermediates that leads to the exclusive presence of myristic acid in glycolipid A and, ultimately, in the variant surface glycoprotein (VSG). In the presence of DFP, the de novo synthesis of GPI precursors cannot proceed beyond glycolipid C' (the unremodelled version of glycolipid C) and lyso-glycolipid C'. Under these conditions glycolipid C'-type GPI anchors appear on newly synthesized VSG molecules. However, the efficiencies of both anchor addition to VSG and N-glycosylation of VSG were significantly reduced. A modified model of the GPI biosynthetic pathway in bloodstream form African trypanosomes incorporating these findings is presented.

MeSH Terms
Acylation/drug effects Animals Fatty Acids/metabolism Glycolipids/metabolism Glycoproteins/metabolism Glycosylation Glycosylphosphatidylinositols/biosynthesis,chemistry Inositol/metabolism Isoflurophate/pharmacology Phenylmethylsulfonyl Fluoride/pharmacology Rats Trypanosoma brucei brucei/drug effects,metabolism Tunicamycin/pharmacology
Chemicals
Fatty Acids Glycolipids Glycoproteins Glycosylphosphatidylinositols Tunicamycin Isoflurophate Inositol Phenylmethylsulfonyl Fluoride
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Güther M L
Department of Biochemistry, University of Dundee, UK.
Ferguson M A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1995-07-03
Pages
3080-93
Language
English
Region
England
NLM ID
8208664
PMCID
PMC394370
Subset
IM
Grants
Wellcome Trust · United Kingdom
External Links
PubMed source PMC Full Text View journal details
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