In situ assay of the intramitochondrial enzymes: use of alamethicin for permeabilization of mitochondria.
The channel-forming antibiotic alamethicin was used to permeabilize mitochondrial membranes for the low molecular mass hydrophilic substrates NADH and ATP. Alamethicin-treated mitochondria show high rotenone-sensitive NADH oxidase, NADH-quinone reductase, and oligomycin-sensitive and carboxyatractylate-insensitive ATPase activities. Alamethicin does not affect Complex I and ATPase activities in inside-out submitochondrial particles. Permeabilized mitochondria quantitatively retain their aconitase and iso-citrate dehydrogenase activities. Electron microscopy of alamethicin-treated mitochondria reveals no disruption of their outer and inner membranes. From the results obtained it is recommended, that alamethicin be used for the in situ catalytic assay of intramitochondrially located enzymes.
Gostimskaya
Irina S
IS
Department of Biochemistry, School of Biology, Moscow State University, Moscow 119992, Russian Federation.
Grivennikova
Vera G
VG
Zharova
Tatyana V
TV
Bakeeva
Lora E
LE
Vinogradov
Andrei D
AD
D000408:Alamethicin / Q000737:chemistry*; D001681:Biological Assay; D008854:Microscopy, Electron; D008928:Mitochondria / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure; D010539:Permeability
D000408:Alamethicin