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J. Biol. Chem., Vol. 266, Issue 23, 15170-15179, Aug, 1991
MJ McConville and JM Blackwell
In addition to utilizing glycosylated phosphatidylinositols (GPIs) as
anchors for surface proteins, protozoan parasites of the genus Leishmania
synthesize two novel classes of GPI: the polydisperse lipophosphoglycans
(LPGs) and a family of low molecular weight glycoinositol phospholipids
(GIPLs). We now show that LPG is expressed in high copy number (6 x 10(6)
molecules/cell) in the promastigote (insect) stage of L. donovani but not
in the amastigote stage, which infects mammalian macrophages. Detection of
these molecules was by gas chromatography-mass spectrometric analyses and
by a sensitive radiolabeling procedure. In contrast, a novel family of
GIPLs was present in high copy number (approximately 10(7) molecules/cell)
in both promastigote and amastigote stages of L. donovani. These
glycolipids were purified and analyzed by gas chromatography-mass
spectrometry, methylation analysis, and by chemical and enzymatic
sequencing after deamination and NaB3H4 reduction. Promastigotes contained
three major GIPLs species with the following generalized structure
[formula: see text] where R = H for isoM2, Man alpha 1- for isoM3 or Man
alpha 1-2Man alpha 1- for isoM4. Amastigotes contained two major GIPL
species that lacked the alpha 1-3-linked mannose branch and had the linear
structures Man alpha 1-6Man alpha 1-4GlcN (M2) and Man alpha 1-2Man alpha
1-6Man alpha 1-4GlcN (M3) linked to alkylacyl-PI. The 1-O-alkyl-2-acyl-PI
moieties of all these species contained predominantly C18:0 alkyl chains
and C16:0 or C18:0 fatty acids. Amastigotes contained, in addition, a
GalNAc beta 1-3 terminating glycosphingolipid with homology to the
mammalian para Forssman glycolipid. This glycolipid appeared to be a
constituent of the parasite membrane but was not metabolically labeled with
[3H]glucose, suggesting that it was acquired from host cells. These results
suggest that LPG may not be required for amastigote survival in the
mammalian host and that the GIPLs are likely to be major components on the
surface membrane in both stages.
Developmental changes in the glycosylated phosphatidylinositols of Leishmania donovani. Characterization of the promastigote and amastigote glycolipids
Department of Biochemistry, University of Dundee, United Kingdom.
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