Hemozoin-free Plasmodium falciparum mitochondria for physiological and drug susceptibility studies

作者：Mather Michael W^*; Morrisey Joanne M; Vaidya Akhil B

来源：Molecular and Biochemical Parasitology, 2010, 174(2): 150-153.

DOI：10.1016/j.molbiopara.2010.07.006

• Cell fractionation
• Mitochondria
• Malaria
• Hemozoin
• Magnetic separation
• Spectrophotometric enzyme assay

Isolation of mitochondria of high purity and with intact enzymatic activities from malaria parasites has proven to be a major obstacle in characterizing the parasite mitochondrial physiology We describe here an improved procedure for the isolation of a mitochondrially enriched preparation from the trophozoite stage of erythrocytic Plasmodium falciparum, combining disruption by N(2) cavitation and differential centrifugation with magnetic removal of hemozoin-associated material These mitochondrial preparations may be used to assay various mitochondrial enzyme activities, such as succinate and dihydroorotate dehydrogenases, ubiquinol-cytochrome c oxidoreductase, and cytochrome c oxidase They also exhibit a low level of ATPase activity, which is only marginally inhibited by classical inhibitors We have used this prepar

• 出版日期2010-12