Purification and characterization of an extracellular phosphoglycan from Leishmania donovani.
An extracellular phosphoglycan (exPG), present in the culture medium of the promastigote form of Leishmania donovani, was purified and structurally characterized. The purification scheme included ethanol precipitation of the culture medium, anion exchange chromatography, hydrophobic chromatography on phenyl-Sepharose, and preparative polyacrylamide gel electrophoresis. Structural analysis by 1H-1H NMR, methylation linkage analysis, and glycosidase digestion revealed that the exPG consisted of the following structure: (CAP)----[PO4-6Galp beta 1-4Manp alpha 1]10-11-PO4-6Galp beta 1-4Man. The cap was found to be one of several small, neutral oligosaccharides, the most abundant of which was the trisaccharide Galp beta 1-4(Manp alpha 1-2)Man. The results indicated structural analogy to the cellular-derived lipophosphoglycan (LPG) from L. donovani. The important exceptions are a lack of the lipid anchor, the entire phosphosaccharide core, and several of the repeating disaccharide units. Although the function of exPG is presently unknown, it may play a protective role for the promastigote in the insect vector or during infection of a mammalian host.
Greis
K D
KD
Department of Biochemistry, University of Kentucky Medical Center, Lexington 40536.
Turco
S J
SJ
Thomas
J R
JR
McConville
M J
MJ
Homans
S W
SW
Ferguson
M A
MA
D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D008401:Gas Chromatography-Mass Spectrometry; D007202:Indicators and Reagents; D007893:Leishmania donovani / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D011134:Polysaccharides / Q000737:chemistry* / Q000302:isolation & purification; D014316:Tritium
D007202:Indicators and Reagents; D009844:Oligosaccharides; D011134:Polysaccharides; D014316:Tritium