Mitochondrial metabolite carrier proteins: purification, reconstitution, and transport studies.

摘要: Publisher Summary This chapter also discusses the methods for purifying metabolite carriers from mitochondria, reconstituting them in liposomes, and their transport activities reconstituted system. The general purification scheme applied isolation of mitochondrial functional state involves solubilization by nonionic detergents; chromatography on hydroxyapatite, recovering respective carrier eluate; final using various further chromatographic procedures. detailed procedures phosphate, oxoglutarate/malate, dicarboxylates, tricarboxylates (citrate), carnitine/acylcarnitines, aspartate/glutamate, ornithine/citrulline functionally active are described. For characterization terms activity, purified proteins have to be into liposomal membranes. selection reconstitution is restricted fact that these denatured detergents with high critical micellar concentration; thus, dialysis related cannot applied.