title: Protein kinase Cepsilon interacts with and inhibits the permeability transition pore in cardiac mitochondria.
abstract: Although functional coupling between protein kinase Cepsilon (PKCepsilon) and mitochondria has been implicated in the genesis of cardioprotection, the signal transduction mechanisms that enable this link and the identities of the mitochondrial proteins modulated by PKCepsilon remain unknown. Based on recent evidence that the mitochondrial permeability transition pore may be involved in ischemia/reperfusion injury, we hypothesized that protein-protein interactions between PKCepsilon and mitochondrial pore components may serve as a signaling mechanism to modulate pore function and thus engender cardioprotection. Coimmunoprecipitation and GST-based affinity pull-down from mouse cardiac mitochondria revealed interaction of PKCepsilon with components of the pore, namely voltage-dependent anion channel (VDAC), adenine nucleotide translocase (ANT), and hexokinase II (HKII). VDAC1, ANT1, and HKII were present in the PKCepsilon complex at approximately 2%, approximately 0.2%, and approximately 1% of their total expression, respectively. Moreover, in vitro studies demonstrated that PKCepsilon can directly bind and phosphorylate VDAC1. Incubation of isolated cardiac mitochondria with recombinant PKCepsilon resulted in a significant inhibition of Ca2+-induced mitochondrial swelling, an index of pore opening. Furthermore, cardiac-specific expression of active PKCepsilon in mice, which is cardioprotective, greatly increased interaction of PKCepsilon with the pore components and inhibited Ca2+-induced pore opening. In contrast, cardiac expression of kinase-inactive PKCepsilon did not affect pore opening. Finally, administration of the pore opener atractyloside significantly attenuated the infarct-sparing effect of PKCepsilon transgenesis. Collectively, these data demonstrate that PKCepsilon forms physical interactions with components of the cardiac mitochondrial pore. This in turn inhibits the pathological function of the pore and contributes to PKCepsilon-induced cardioprotection.
authors.affiliation: Department of Physiology and Biophysics, University of Louisville, Louisville, Ky, USA. CPBaines@gmx.net
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Baines
authors.forename: Chang-Xu
authors.initials: CX
authors.lastname: Song
authors.forename: Yu-Ting
authors.initials: YT
authors.lastname: Zheng
authors.forename: Guang-Wu
authors.initials: GW
authors.lastname: Wang
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Ou-Li
authors.initials: OL
authors.lastname: Wang
authors.forename: Yiru
authors.initials: Y
authors.lastname: Guo
authors.forename: Roberto
authors.initials: R
authors.lastname: Bolli
authors.forename: Ernest M
authors.initials: EM
authors.lastname: Cardwell
authors.forename: Peipei
authors.initials: P
authors.lastname: Ping
mesh_terms: D000818:Animals; D001278:Atractyloside / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006593:Hexokinase / Q000235:genetics / Q000378:metabolism; D015151:Immunoblotting; D007425:Intracellular Membranes / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D008929:Mitochondria, Heart / Q000187:drug effects / Q000378:metabolism*; D000226:Mitochondrial ADP, ATP Translocases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D009203:Myocardial Infarction / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D015428:Myocardial Reperfusion Injury / Q000150:complications; D010539:Permeability; D010766:Phosphorylation; D018272:Porins / Q000235:genetics / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D051744:Protein Kinase C-epsilon; D051381:Rats; D050995:Voltage-Dependent Anion Channel 1; D050994:Voltage-Dependent Anion Channels
chemical_list: D004791:Enzyme Inhibitors; D018272:Porins; C494763:Vdac1 protein, mouse; C494764:Vdac1 protein, rat; D050994:Voltage-Dependent Anion Channels; D001278:Atractyloside; D000226:Mitochondrial ADP, ATP Translocases; D050995:Voltage-Dependent Anion Channel 1; C497256:Prkce protein, mouse; C497257:Prkce protein, rat; D006593:Hexokinase; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon