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Mol Biochem Parasitol. 1981 Dec 31;4(5-6):243-53.

Growth of human malaria parasites in biotinylated erythrocytes.

Abstract

The adaptation of the biotin-avidin system for the analysis of membrane pathobiology in Plasmodium falciparum malaria is described. Biotin was linked covalently via the succinimide ester derivative (biotinyl-N-hydroxysuccinimide ester, BNHS) to intact human erythrocytes, prior to inoculation and in vitro cultivation of falciparum parasites. Growth experiments indicated that incubation concentrations of less than 1.0 mg BNHS/1.0 ml erythrocyte packed cell volume could yield biotinylated erythrocytes capable of sustaining parasite growth at levels comparable to control cultures. Using a synthesized [14C]BNHS compound at optimal incubation concentration, it was determined that 1.32 X 10(-4) mmol [14C]BNHS were bound per 1.0 mg of erythrocyte stromal protein. In addition, analysis of [14C]biotinylated red blood cell ghost preparations by polyacrylamide gel electrophoresis demonstrated that band 3 (a heterogeneous glycoprotein) was the principal site of membrane labeling. Approximately 77% of total membrane-associated [14C]BNHS was localized to this polypeptide. The unique properties of the specific, ligand-protein interaction of the biotin-avidin complex suggest that the biotinylation procedure described in this report will provide a useful analytical tool in host cell-plasmodial parasite, membrane studies.

PMID:
7038477
[Indexed for MEDLINE]

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