Cyclophilin D modulates mitochondrial F0F1-ATP synthase by interacting with the lateral stalk of the complex.
Blue native gel electrophoresis purification and immunoprecipitation of F(0)F(1)-ATP synthase from bovine heart mitochondria revealed that cyclophilin (CyP) D associates to the complex. Treatment of intact mitochondria with the membrane-permeable bifunctional reagent dimethyl 3,3-dithiobis-propionimidate (DTBP) cross-linked CyPD with the lateral stalk of ATP synthase, whereas no interactions with F(1) sector subunits, the ATP synthase natural inhibitor protein IF1, and the ATP/ADP carrier were observed. The ATP synthase-CyPD interactions have functional consequences on enzyme catalysis and are modulated by phosphate (increased CyPD binding and decreased enzyme activity) and cyclosporin (Cs) A (decreased CyPD binding and increased enzyme activity). Treatment of MgATP submitochondrial particles or intact mitochondria with CsA displaced CyPD from membranes and activated both hydrolysis and synthesis of ATP sustained by the enzyme. No effect of CsA was detected in CyPD-null mitochondria, which displayed a higher specific activity of the ATP synthase than wild-type mitochondria. Modulation by CyPD binding appears to be independent of IF1, whose association to ATP synthase was not affected by CsA treatment. These findings demonstrate that CyPD association to the lateral stalk of ATP synthase modulates the activity of the complex.
Giorgio
Valentina
V
Department of Biomedical Sciences and the Consiglio Nazionale delle Ricerche Institute of Neuroscience, University of Padova, I-35121 Padova, Italy.
Bisetto
Elena
E
Soriano
Maria Eugenia
ME
Dabbeni-Sala
Federica
F
Basso
Emy
E
Petronilli
Valeria
V
Forte
Michael A
MA
Bernardi
Paolo
P
Lippe
Giovanna
G
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D003510:Cyclohexanes / Q000494:pharmacology; D000081406:Peptidyl-Prolyl Isomerase F; D021983:Cyclophilins / Q000378:metabolism / Q000502:physiology*; D006321:Heart / Q000502:physiology; D047468:Immunoprecipitation; D051379:Mice; D008929:Mitochondria, Heart / Q000201:enzymology* / Q000378:metabolism; D008930:Mitochondria, Liver / Q000201:enzymology* / Q000378:metabolism; D025261:Mitochondrial Proton-Translocating ATPases / Q000378:metabolism*; D008954:Models, Biological; D011485:Protein Binding
D003510:Cyclohexanes; D000081406:Peptidyl-Prolyl Isomerase F; C000631446:PPIF protein, mouse; C067220:di-tert-butylperoxy-3,3,5-trimethylcyclohexane peroxide; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D025261:Mitochondrial Proton-Translocating ATPases; D021983:Cyclophilins