Isolation and analysis of nucleotides from erythrocyte-free malarial parasites (Plasmodium berghei) and potential relevance to malaria chemotherapy

K. Van Dyke, M. A. Trush, M. E. Wilson, P. K. Stealey

Research output: Contribution to journalArticlepeer-review

7 Scopus citations


Studies using erythrocyte free preparations of P. berghei were conducted with a view to improving knowledge of parasite metabolism, particularly nucleotide metabolism. The free parasites employed in these studies were prepared by saponin lysis of parasitized mouse erythrocytes in isotonic glucose solutions. A comparative study of post lytic metabolic activity of free parasites prepared by saponin, ammonium chloride, or osmotic lysis indicated a significantly greater retention of metabolic activity in the saponin lysis preparations. Separations of nucleoside mono, di, and triphosphates extracted from free parasites were performed by means of high pressure liquid chromatography (HPLC), and ATP was additionally measured by luciferin luciferase assay. Studies designed to differentiate among uptake, phosphorylation, and subsequent incorporation of 3H adenosine into nucleic acids of the free parasite strongly suggested that adenosine is metabolized either outside or on the parasite membrane, being first deaminated to inosine and then deribosylated to hypoxanthine. Observations from HPLC and radioisotope precursor studies support a hypothesis in which hypoxanthine may be proposed as being a pivotal substrate for purine salvage by malarial parasites. Some of the key steps in purine salvage and pyrimidine biosynthesis were investigated, using radiolabel uptake studies and HPLC analysis of nucleotides of the free malarial parasite. These studies suggest that hypoxanthine uptake may constitute an important new basis for chemotherapeutic attack on the malarial parasite.

Original languageEnglish (US)
Pages (from-to)253-264
Number of pages12
JournalBulletin of the World Health Organization
Issue number2-3
StatePublished - Dec 1 1977

ASJC Scopus subject areas

  • Public Health, Environmental and Occupational Health


Dive into the research topics of 'Isolation and analysis of nucleotides from erythrocyte-free malarial parasites (Plasmodium berghei) and potential relevance to malaria chemotherapy'. Together they form a unique fingerprint.

Cite this