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Labeling and initial characterization of polar lipids in cultures ofPlasmodium falciparum

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Abstract

The present report describes the radioactive labeling of polar lipids in in vitro cultures ofPlasmodium falciparum as well as their extraction with organic solvents and their partial characterization by chemical and enzymatic methods. All substances detected could be cleaved by alkali, suggesting that they were esters rather than sphingolipids or compounds containing alkyl groups. Dolichol-cycle intermediates were not detected. Phosphatidylinositol, phosphatidylethanolamine, and phosphatidylcholine were labeled by fatty acids and inositol or ethanolamine, respectively, confirming their de novo synthesis by the parasite. Metabolic labeling with glucosamine and cleavage by phosphatidylinositol-specific phospholipase C provided evidence of the formation ofN-acetyl-glucosaminylphosphatidylinositol, an obligate precursor in the biosynthesis of glycosylphosphatidylinositol membrane anchors of proteins.

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Abbreviations

Cho :

Choline

EDTA :

ethylenediaminetetraacetic acid

EGTA :

ethyleneglycol-bis-(β-aminoether)-N,N′-tetraacetic acid

EtN :

ethanolamine

Fuc :

fucose

Gal :

galactose

Glc :

glucose

GlcN :

glucosamine

GlcNAc :

N-acetyl glucosamine

HEPES:

N-2-hydroxyethylpiperazinyl-N′-2-ethanesulfonic acid

HPAEC :

high pH anion-exchange chromatography

Man :

mannose

NP-40:

Nonidet P-40

PI:

phosphatidylinositol

PE :

phosphatidylethanolamine

PCho :

phosphatidylcholine

PMSF :

phenylmethylsulfonyl fluoride

TLC :

thin-layer chromatography

TLCK :

tosyllysinechloromethylketone

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Authors and Affiliations

  1. Zentrum für Hygiene und Medizinische Mikrobiologie, Universität Marburg, Robert-Koch-Str. 17, W-3550, Marburg, Germany

    A. Dieckmann-Schuppert, S. Bender & R. T. Schwarz

  2. Division of Parasitology, National Institute for Medical Research, NW7 1AA, London

    A. A. Holder

  3. Dept. of Microbiology and Immunology, Stanford University, 94305, Stanford, CA, USA

    K. Haldar

Authors
  1. A. Dieckmann-Schuppert
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  2. S. Bender
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  3. A. A. Holder
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  4. K. Haldar
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  5. R. T. Schwarz
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Additional information

This study was supported by grants Schw 296/4-1 and 296/4-2 from the Deutsche Forschungsgemeinschaft, by the British-German Academic Research Collaboration (ARC) Program of the German Academic Exchange Service (DAAD), by the Fonds der Chemischen Industrie, by the Hessisches Ministerium für Wissenschaft und Kunst, and by the P.E. Kempes Foundation, Marburg

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Dieckmann-Schuppert, A., Bender, S., Holder, A.A. et al. Labeling and initial characterization of polar lipids in cultures ofPlasmodium falciparum . Parasitol Res 78, 416–422 (1992). https://doi.org/10.1007/BF00931698

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  • DOI: https://doi.org/10.1007/BF00931698

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