Abstract
Treatment of rat heart mitochondria with phosphate or mersalyl releases a number of proteins, including the mitochondrial creatine kinase (mt-CK). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the released proteins showed that phosphate is more selective than mersalyl in releasing mt-CK. The rebinding of mt-CK to mitochondria was selectively inhibited by adriamycin, which complexes membrane-bound cardiolipin. mt-CK activity and binding experiments have shown that intact mitochondria are able to bind approximately twice the amount of mt-CK they originally contain. Liver mitochondria bound heart mitochondria mt-CK to the same extent as creatine kinase-depleted heart mitochondria. mt-CK was bound by liposomes but only if they contained cardiolipin. The binding of mt-CK to cardiolipin-containing liposomes was inhibited by adriamycin. Phosphatidylcholine liposomes reconstituted with the purified ADP/ATP translocator failed to bind mt-CK.
Highlights
Treatment of rat heart mitochondriawith phosphate elimination of the releasing agenthas been shown to be or mersalyl releases a number of proteins, including inhibited by adriamycin, a drug widely used in cancer chemthe mitochondrial creatinekinase
The resulting suspenmt-CK’ was discovered by Jacobs et al [1]in muscle and brain mitochondria.Heart mitochondria possess a significant amount of mitochondrial creatinekinase (mt-CK), but liver and kidney mitochondria don’t [1]. mt-CK is a water-soluble enzyme bound tothe sion was centrifuged for 60 min a t 150,000X g, and the pellet was resuspended inbuffer B
Release and Binding of mt-CK in Heart and Liver Mitochondria-The proteins released by mersalyl or Pi treatment were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (Fig. 1, lanes 3 and 5, respectively)
Summary
From the Laboratory of Biochemistry, Swiss Federal Instituteof Technology (ETH), CH-8092 Zurich, Switzerland. Treatment of rat heart mitochondriawith phosphate elimination of the releasing agenthas been shown to be or mersalyl releases a number of proteins, including inhibited by adriamycin, a drug widely used in cancer chemthe mitochondrial creatinekinase (mt-CK). Sodium otherapy [10].Adriamycin has been shown to bind dodecyl sulfate-polyacrylamide gel electrophoresis of to cardiolipin in the inner mitochondial membrane [11]and the released proteins showed that phosphate is more to inhibit the activity of cardiolipin-dependent proteins like selective than mersalyl in releasing mt-CK. The rebinding of mt-CK to mitochondria was selectively inhibited by adriamycin, which complexes membranebound cardiolipin. In this work evidence will be presented that mt-CK binds to cardiolipin associated with the inner membrane in a reaction which is inhibited by adriamycin. Liver mitochondria bound heart mitochondria mt-CK to the same extent as creatine
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