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Submitted on August 9, 2002
Revised on March 12, 2003
Accepted on March 18, 2003
Interacts With and Inhibits the Permeability Transition Pore in Cardiac Mitochondria
From the Departments of Physiology and Biophysics and Medicine/Division of Cardiology (C.P.B., C.-X.S., Y.-T.Z., O.-L.W., Y.G., R.B.), University of Louisville, Louisville, Ky; Departments of Physiology and Medicine/Division of Cardiology (G.-W.W., J.Z., E.M.C., P.P.), University of California at Los Angeles, Los Angeles, Calif.
* To whom correspondence should be addressed. E-mail: CPBaines{at}gmx.net.
Although functional coupling between protein kinase C
(PKC
) 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 PKC
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 PKC
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 PKC
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 PKC
complex at
2%,
0.2%, and
1% of their total expression, respectively. Moreover, in vitro studies demonstrated that PKC
can directly bind and phosphorylate VDACs. Incubation of isolated cardiac mitochondria with recombinant PKC
resulted in a significant inhibition of Ca2+-induced mitochondrial swelling, an index of pore opening. Furthermore, cardiac-specific expression of active PKC
in mice, which is cardioprotective, greatly increased interaction of PKC
with the pore components and inhibited Ca2+-induced pore opening. In contrast, cardiac expression of kinase-inactive PKC
did not affect pore opening. Finally, administration of the pore opener atractyloside significantly attenuated the infarct-sparing effect of PKC
transgenesis. Collectively, these data demonstrate that PKC
forms physical interactions with components of the cardiac mitochondrial pore. This in turn inhibits the pathological function of the pore and contributes to PKC
-induced cardioprotection.
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