Inhibition of the mitochondrial permeability transition by creatine kinase substrates. Requirement for microcompartmentation.
Mitochondria from transgenic mice, expressing enzymatically active mitochondrial creatine kinase in liver, were analyzed for opening of the permeability transition pore in the absence and presence of creatine kinase substrates but with no external adenine nucleotides added. In mitochondria from these transgenic mice, cyclosporin A-inhibited pore opening was delayed by creatine or cyclocreatine but not by beta-guanidinopropionic acid. This observation correlated with the ability of these substrates to stimulate state 3 respiration in the presence of extramitochondrial ATP. The dependence of transition pore opening on calcium and magnesium concentration was studied in the presence and absence of creatine. If mitochondrial creatine kinase activity decreased (i.e. by omitting magnesium from the medium), protection of permeability transition pore opening by creatine or cyclocreatine was no longer seen. Likewise, when creatine kinase was added externally to liver mitochondria from wild-type mice that do not express mitochondrial creatine kinase in liver, no protective effect on pore opening by creatine and its analog was observed. All these findings indicate that mitochondrial creatine kinase activity located within the intermembrane and intercristae space, in conjunction with its tight functional coupling to oxidative phosphorylation, via the adenine nucleotide translocase, can modulate mitochondrial permeability transition in the presence of creatine. These results are of relevance for the design of creatine analogs for cell protection as potential adjuvant therapeutic tools against neurodegenerative diseases.
Dolder
Max
M
Institute of Cell Biology, Swiss Federal Institute of Technology, ETH-Hönggerberg, CH-8093 Zürich, Switzerland. dolder@biotech.biol.ethz.ch
Walzel
Bernd
B
Speer
Oliver
O
Schlattner
Uwe
U
Wallimann
Theo
T
D000225:Adenine / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002470:Cell Survival; D002855:Chromatography, Thin Layer; D003401:Creatine / Q000494:pharmacology; D003402:Creatine Kinase / Q000737:chemistry* / Q000378:metabolism; D003404:Creatinine / Q000031:analogs & derivatives* / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006146:Guanidines / Q000378:metabolism; D007473:Ion Channels / Q000037:antagonists & inhibitors*; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008274:Magnesium / Q000378:metabolism / Q000494:pharmacology; D051379:Mice; D008822:Mice, Transgenic; D008928:Mitochondria / Q000378:metabolism; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D008954:Models, Biological; D019636:Neurodegenerative Diseases / Q000473:pathology; D010100:Oxygen / Q000378:metabolism; D010101:Oxygen Consumption; D010725:Phosphocreatine / Q000378:metabolism; D010766:Phosphorylation; D011422:Propionates / Q000378:metabolism; D011485:Protein Binding; D013997:Time Factors
D006146:Guanidines; D007473:Ion Channels; D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D011422:Propionates; D010725:Phosphocreatine; D000244:Adenosine Diphosphate; C012260:cyclocreatine; D000255:Adenosine Triphosphate; D003404:Creatinine; D003402:Creatine Kinase; D008274:Magnesium; D000225:Adenine; D003401:Creatine; D010100:Oxygen; D002118:Calcium; C005775:guanidinopropionic acid