Solubilization and partial purification of glycerol-3-phosphate oxidase from mitochondria of Trypanosoma brucei

Trypanosomes are flagellated protozoa responsible for several serious parasitic diseases of humans and domestic animals. Bloodstream forms of Trypanosoma brucei and T. rhodesiense lack cytochromes, are completely dependent on glycolysis for energy, and respiration in whole cells is insensitive to inhibitors such as cyanide, azide, and antimycin A. Studies by several investigators have shown that bloodstream forms have a KCN-insensitive L- alpha -glycerophosphate oxidase, which consists of two components: a flavin-linked glycerol-3-phosphate (GPO) dehydrogenase (EC 1.1.99.5) and an oxidase which are probably linked via ubiquinol. With the use of a fast pressure liquid chromatography (FPLC) system and a Mono Q anion-exchange column, we have been able to solubilize and partially purify the GPO activity.