Plasmodium berghei: High pressure liquid chromatographic analysis of nucleotides from erythrocyte-free malarial parasites

Knox Van Dyke, Mark Wilson, Michael Trush, Michael Taylor, Patricia Stealey

Research output: Contribution to journalArticle

Abstract

Isolation and analysis of the nucleotides from the malarial parasite, Plasmodium berghei, are described. This was accomplished by reducing contaminating leukocytes and platelets by settling in 6% dextran-Krebs glucose. Contaminating red cells (either parasitized or nonparasitized) were lysed by a technique using a dilute saponin solution in a Krebs-glucose medium. The parasites were collected by centrifugation and destroyed by addition of 10% trichloroacetic acid. The acid was extracted with Alamine-Freon TF and the nucleotides analyzed using high pressure liquid chromatography with microparticle anion exchange columns. The following compounds were separated and quantitated: ATP, GTP, CTP, UTP, ADP, GDP, CDP, UDP, AMP, GMP, CMP, UMP, and NAD. In nanomoles of nucleoside triphosphate isolated, the order of highest to lowest concentration was: ATP > UTP > GTP > CTP, while the order of diphosphates was: ADP > CDP > UDP > GDP, and the monophosphates: NAD > AMP > CMP > UMP > GMP. The order in amount of nucleotides present was: nucleoside triphosphates > nucleoside monophosphates > nucleoside diphosphates. The statistical analysis of the nucleoside triphosphates showed little deviation from sample to sample. A comparison of the sample from the host mouse red blood cell shows significant concentrations only of ATP, ADP, and NAD, indicating that contamination from host blood cells was unlikely.

Original languageEnglish (US)
Pages (from-to)274-281
Number of pages8
JournalExperimental Parasitology
Volume42
Issue number2
DOIs
StatePublished - Aug 1977

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Keywords

  • High pressure liquid chromatography
  • Malaria
  • Mouse
  • Nucleotides
  • Parasitic protozoa
  • Plasmodium berghei
  • Purine/pyrimidine metabolism

ASJC Scopus subject areas

  • Parasitology
  • Immunology
  • Infectious Diseases

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