Study on heart mitochondrial creatine kinase using a cross-linking bifunctional reagent. I. The binding involves the octameric form of the enzyme.
Bovine heart mitochondria suspended in 0.25 M sucrose were treated with 0.3% glutaraldehyde (GA). The membranes were disintegrated by ultrasonication in 0.25 M KCl and precipitated by centrifugation. The supernatant was assayed for creatine kinase (CKm) oligomeric forms by ultracentrifugation in a sucrose density gradient. A kinetic analysis of membrane-bound CKm was performed before and after ultrasonication. The data obtained suggest that the CKm octamer is the only form of CKm bound to mitochondrial membranes during GA treatment. This finding was confirmed by an analysis of extracts from untreated mitochondria using high resolution gel filtration.
Lipskaya TYu
Department of Biochemistry, School of Biology, M.V. Lomonosov Moscow State University, USSR.
Trofimova
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D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D003402:Creatine Kinase / Q000032:analysis*; D003432:Cross-Linking Reagents; D005976:Glutaral / Q000302:isolation & purification; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology*; D011506:Proteins / Q000032:analysis; D013385:Succinate Dehydrogenase / Q000378:metabolism; D014465:Ultrasonics
D003432:Cross-Linking Reagents; D011506:Proteins; D013385:Succinate Dehydrogenase; D003402:Creatine Kinase; D005976:Glutaral