Conditioning the heart induces formation of signalosomes that interact with mitochondria to open mitoKATP channels.
Perfusion of the heart with bradykinin triggers cellular signaling events that ultimately cause opening of mitochondrial ATP-sensitive K+ (mitoKATP) channels, increased H2O2 production, inhibition of the mitochondrial permeability transition (MPT), and cardioprotection. We hypothesized that the interaction of bradykinin with its receptor induces the assembly of a caveolar signaling platform (signalosome) that contains the enzymes of the signaling pathway and that migrates to mitochondria to induce mitoKATP channel opening. We developed a novel method for isolating and purifying signalosomes from Langendorff-perfused rat hearts treated with bradykinin. Fractions containing the signalosomes were found to open mitoKATP channels in mitochondria isolated from untreated hearts via the activation of mitochondrial PKC-epsilon. mitoKATP channel opening required signalosome-dependent phosphorylation of an outer membrane protein. Immunodetection analysis revealed the presence of the bradykinin B2 receptor only in the fraction isolated from bradykinin-treated hearts. Immunodetection and immunogold labeling of caveolin-3, as well as sensitivity to cholesterol depletion and resistance to Triton X-100, attested to the caveolar nature of the signalosomes. Ischemic preconditioning, ischemic postconditioning, and perfusion with ouabain also led to active signalosome fractions that opened mitoKATP channels in mitochondria from untreated hearts. These results provide initial support for a novel mechanism for signal transmission from a plasma membrane receptor to mitoKATP channels.
Quinlan
Casey L
CL
Department of Biology, Portland State University, PO Box 751, Portland, OR 97201-0751, USA.
Costa
Alexandre D T
AD
Costa
Cinthia L
CL
Pierre
Sandrine V
SV
Dos Santos
Pierre
P
Garlid
Keith D
KD
D000465:Algorithms; D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D002316:Cardiotonic Agents / Q000494:pharmacology; D017869:Cyclic GMP-Dependent Protein Kinases / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015151:Immunoblotting; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D019157:Ischemic Preconditioning, Myocardial; D054086:KATP Channels / Q000187:drug effects*; D018942:Macrolides / Q000494:pharmacology; D008297:Male; D008929:Mitochondria, Heart / Q000502:physiology*; D017830:Octoxynol / Q000494:pharmacology; D010042:Ouabain / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D051744:Protein Kinase C-epsilon / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000502:physiology*; D047392:beta-Cyclodextrins / Q000494:pharmacology
D002316:Cardiotonic Agents; D004791:Enzyme Inhibitors; D054086:KATP Channels; D018942:Macrolides; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; D010042:Ouabain; C040929:bafilomycin A1; D017830:Octoxynol; D017869:Cyclic GMP-Dependent Protein Kinases; D051744:Protein Kinase C-epsilon; D001920:Bradykinin