March 22, 2006Journal of Biological Chemistry131 citationsOpen Access

Suppression of Apoptosis by Cyclophilin D via Stabilization of Hexokinase II Mitochondrial Binding in Cancer Cells

KMKiyotaka MachidaWaseda University YOYoshihiro OhtaKawasaki Medical School HOHiroyuki OsadaMicrobial Chemistry Research Foundation

Key Points

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Abstract

The permeability transition pore is involved in the mitochondrial pathway of apoptosis. Cyclophilin D, a pore component, has catalytic activity as a peptidyl prolyl cis, trans-isomerase (PPIase), which is essential to the pore opening. It has been reported that cyclophilin D overexpression suppresses apoptosis in cancer cells. To clarify the mechanism of this effect, we generated glioma cells overexpressing wild-type or a PPIase-deficient mutant of cyclophilin D. Interestingly, we found that the PPIase-dependent apoptosis suppression by cyclophilin D correlated with the amounts of mitochondrial-bound hexokinase II, which has anti-apoptotic activity. Inactivation of endogenous cyclophilin D by small interference RNA or a cyclophilin inhibitor was found to release hexokinase II from mitochondria and to enhance Bax-mediated apoptosis. The anti-apoptotic effects of cyclophilin D were canceled out by the detachment of hexokinase II from mitochondria, demonstrating that mitochondrial binding of hexokinase II is essential to the apoptosis suppression by cyclophilin D. Furthermore, cyclophilin D dysfunction appears to abrogate hexokinase II-mediated apoptosis suppression, indicating that cyclophilin D is required for the anti-apoptotic activity of hexokinase II. Based on the above, we propose here that cyclophilin D suppresses apoptotic cell death via a mitochondrial hexokinase II-dependent mechanism in cancer cells. The permeability transition pore is involved in the mitochondrial pathway of apoptosis. Cyclophilin D, a pore component, has catalytic activity as a peptidyl prolyl cis, trans-isomerase (PPIase), which is essential to the pore opening. It has been reported that cyclophilin D overexpression suppresses apoptosis in cancer cells. To clarify the mechanism of this effect, we generated glioma cells overexpressing wild-type or a PPIase-deficient mutant of cyclophilin D. Interestingly, we found that the PPIase-dependent apoptosis suppression by cyclophilin D correlated with the amounts of mitochondrial-bound hexokinase II, which has anti-apoptotic activity. Inactivation of endogenous cyclophilin D by small interference RNA or a cyclophilin inhibitor was found to release hexokinase II from mitochondria and to enhance Bax-mediated apoptosis. The anti-apoptotic effects of cyclophilin D were canceled out by the detachment of hexokinase II from mitochondria, demonstrating that mitochondrial binding of hexokinase II is essential to the apoptosis suppression by cyclophilin D. Furthermore, cyclophilin D dysfunction appears to abrogate hexokinase II-mediated apoptosis suppression, indicating that cyclophilin D is required for the anti-apoptotic activity of hexokinase II. Based on the above, we propose here that cyclophilin D suppresses apoptotic cell death via a mitochondrial hexokinase II-dependent mechanism in cancer cells. During apoptosis, the permeability of the mitochondrial outer membrane is up-regulated, inducing a release of pro-apoptotic factors such as cytochrome c from the intermembrane space into the cytosol. Once cytochrome c is released from mitochondria, it binds to Apaf-1 and promotes the assembly of apoptosome multiprotein complex, which induces activation of the caspase-9 cell death protease (1Li P. Nijhawan D. Budihardjo I. Srinivasula S.M. Ahmad M. Alnemri E.S. Wang X. Cell. 1997; 91: 479-489Abstract Full Text Full Text PDF PubMed Scopus (6261) Google Scholar). It has been believed that a mitochondrial protein complex called the permeability transition (PT) 2The abbreviations used are: PT, permeability transition; VDAC, voltage-dependent anion channel; ANT, adenine nucleotide translocase; PPIase, peptidyl prolyl cis, trans-isomerase; CTZ, clotrimazole; CsA, cyclosporin A; siRNA, small interference RNA; HA, hemagglutinin; PIPES, 1,4-piperazinediethanesulfonic acid; CHAPS, 3-(3-cholamidopropyl)dimethylammonio-1-propanesulfonic acid. pore, which is composed of the voltage-dependent anion channel (VDAC), the adenine nucleotide translocase (ANT), cyclophilin D, and hexokinase, mediates the permeabilization of the mitochondrial membrane and cytochrome c release (2Beutner G. Rück A. Riede B. Brdiczka D. Biochim. Biophys. Acta. 1998; 1368: 7-18Crossref PubMed Scopus (303) Google Scholar). Cyclophilin D, which is a peptidyl prolyl-cis, trans-isomerase (PPIase), has been considered to be a promoter of pore opening. It has been reported that cyclophilin inhibitors such as cyclosporin A (3Halestrap A.P. Davidson A.M. Biochem. J. 1990; 268: 153-160Crossref PubMed Scopus (672) Google Scholar) and sanglifehrin A (4Clarke S.J. McStay G.P. Halestrap A.P. J. Biol. Chem. 2002; 277: 34793-34799Abstract Full Text Full Text PDF PubMed Scopus (319) Google Scholar) block the opening of the PT pore. Moreover, recent reports clearly show that mitochondria isolated from cyclophilin D null mice are resistant to the permeability transition (5Nakagawa T. Shimizu S. Watanabe T. Yamaguchi O. Otsu K. Yamagata H. Inohara H. Kubo T. Tsujimoto Y. Nature. 2005; 434: 652-657Crossref PubMed Scopus (1372) Google Scholar, 6Baines C.P. Kaiser R.A. Purcell N.H. Blair N.S. Osinska H. Hambleton M.A. Brunskill E.W. Sayen M.R. Gottlieb R.A. Dorn II, G.W. Robbins J. Molkentin J.D. Nature. 2005; 434: 658-662Crossref PubMed Scopus (1866) Google Scholar, 7Basso E. Fante L. Fowlkes J. Petronilli V. Forte M.A. Bernardi P. J. Biol. Chem. 2005; 280: 18558-18561Abstract Full Text Full Text PDF PubMed Scopus (684) Google Scholar, 8Schinzel A.C. Takeuchi O. Huang Z. Fisher J.K. Zhou Z. Rubens J. Hetz C. Danial N.N. Moskowitz M.A. Korsmeyer S.J. Proc. Natl. Acad. Sci. U. S. A. 2005; 102: 12005-12010Crossref PubMed Scopus (713) Google Scholar). Thus, cyclophilin D is now regarded as a key factor in the regulation of pore function. When the PT pore opens in vitro, mitochondria swell, the outer membrane is disrupted, and cytochrome c is released (9Ravagnan L. Marzo I. Costantini P. Susin S.A. Zamzami N. Petit P.X. Hirsch F. Goulbern M. Poupon M.F. Miccoli L. Xie Z. Reed J.C. Kroemer G. Oncogene. 1999; 18: 2537-2546Crossref PubMed Scopus (189) Google Scholar, 10Tafani M. Karpinich N.O. Hurster K.A. Pastorino J.G. Schneider T. Russo M.A. Farber J.L. J. Biol. Chem. 2002; 277: 10073-10082Abstract Full Text Full Text PDF PubMed Scopus (105) Google Scholar). However, during apoptosis cytochrome c release is observed without the permeability transition (11Shimizu S. Tsujimoto Y. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 577-582Crossref PubMed Scopus (265) Google Scholar, 12Scorrano L. Ashiya M. Buttle K. Weiler S. Oakes S.A. Mannella C.A. Korsmeyer S.J. Dev. Cell. 2002; 2: 55-67Abstract Full Text Full Text PDF PubMed Scopus (888) Google Scholar). Indeed, consistent with these observations, cyclophilin D null fibroblasts are not protected from Bax-dependent apoptosis (5Nakagawa T. Shimizu S. Watanabe T. Yamaguchi O. Otsu K. Yamagata H. Inohara H. Kubo T. Tsujimoto Y. Nature. 2005; 434: 652-657Crossref PubMed Scopus (1372) Google Scholar, 6Baines C.P. Kaiser R.A. Purcell N.H. Blair N.S. Osinska H. Hambleton M.A. Brunskill E.W. Sayen M.R. Gottlieb R.A. Dorn II, G.W. Robbins J. Molkentin J.D. Nature. 2005; 434: 658-662Crossref PubMed Scopus (1866) Google Scholar). Therefore, the opening of the PT pore appears to not be essential to Bax-induced apoptosis. However, although the opening of the PT pore is not a trigger of apoptosis, components of the pore are involved in apoptosis as targets or suppressors against pro-apoptotic factors. For instance, VDAC, which is a primary component of the pore, is recognized as a target of pro-apoptotic protein Bax (13Shimizu S. Narita M. Tsujimoto Y. Nature. 1999; 399: 483-487Crossref PubMed Scopus (1928) Google Scholar). VDAC may play a role in regulating cytochrome c release by forming, together with Bax, a highly conductive channel (14Shimizu S. Matsuoka Y. Shinohara Y. Yoneda Y. Tsujimoto Y. J. Cell Biol. 2001; 152: 237-250Crossref PubMed Scopus (332) Google Scholar). It has also been reported that Bax promotes cytochrome c release and apoptosis through interactions with the ANT (15Marzo I. Brenner C. Zamzami N. Jürgensmeier J.M. Susin S.A. Vieira H.L. Prévost M.C. Xie Z. Matsuyama S. Reed J.C. Kroemer G. Science. 1998; 281: 2027-2031Crossref PubMed Scopus (1058) Google Scholar). Hexokinase II, which catalyzes glucose phosphorylation, is overexpressed in cancer cells and plays a pivotal role in cellular and S. A. Biochim. Biophys. Acta. 2002; PubMed Scopus Google Scholar). Hexokinase II appears to the mitochondrial of Bax by the Bax and VDAC J.G. N. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar). Therefore, the mitochondrial binding of hexokinase II to play essential role in the anti-apoptotic mechanism of cancer cells. It has been reported that cyclophilin D suppresses apoptosis it is overexpressed J.D. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar, A. S. PubMed Scopus Google Scholar, Y. N. M. M. M. Biochem. J. PubMed Scopus Google Scholar). However, activity is essential as is the mechanism of the anti-apoptotic of cyclophilin D. the we the of cyclophilin D overexpression or dysfunction on of the PT pore found that cyclophilin D hexokinase II mitochondrial binding by activity. that hexokinase II mitochondrial binding is essential to apoptosis suppression by cyclophilin D. A and were from a protease inhibitor and were from and were from was from Cell and the of Cell and cells were in with of and of in a of cell cyclophilin D were by of the cells were to of cells were for and were The protein of D was by Bax-induced cells of a were for and the cells were with or the cells were with or without CsA, CTZ, and for were and activity was were in and the of in was of were in for The protease activity of was as 2000; PubMed Google Scholar). to release was with a are as the of was a of by the A of was considered c and Bax c release and Bax to mitochondria during apoptosis were as S. J. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar). cells were in the of and for The cells were for and were by c or RNA RNA with to cyclophilin D was from The RNA were as For of endogenous cyclophilin D, we also used RNA that binds the of cyclophilin D The RNA were as Bax was by from The RNA were as A was used as a was into cells with and were isolated from or cells as J.G. N. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar). cells were by with in and of protease inhibitor Cell was by for The was for of mitochondria and were and used for the against cytochrome c and were from was from against against ANT Hexokinase II Bax and against were from and D were from A were from Cell were on with for and with for The cells were with primary and for cells were with of with and from were with a and with and D, we the the (1Li P. Nijhawan D. Budihardjo I. Srinivasula S.M. Ahmad M. Alnemri E.S. Wang X. Cell. 1997; 91: 479-489Abstract Full Text Full Text PDF PubMed Scopus (6261) Google Scholar, G. Rück A. Riede B. Brdiczka D. Biochim. Biophys. Acta. 1998; 1368: 7-18Crossref PubMed Scopus (303) Google Scholar, A.P. Davidson A.M. Biochem. J. 1990; 268: 153-160Crossref PubMed Scopus (672) Google Scholar, S.J. McStay G.P. Halestrap A.P. J. Biol. Chem. 2002; 277: 34793-34799Abstract Full Text Full Text PDF PubMed Scopus (319) Google Scholar, T. Shimizu S. Watanabe T. Yamaguchi O. Otsu K. Yamagata H. Inohara H. Kubo T. Tsujimoto Y. Nature. 2005; 434: 652-657Crossref PubMed Scopus (1372) Google Scholar, 6Baines C.P. Kaiser R.A. Purcell N.H. Blair N.S. Osinska H. Hambleton M.A. Brunskill E.W. Sayen M.R. Gottlieb R.A. Dorn II, G.W. Robbins J. Molkentin J.D. Nature. 2005; 434: 658-662Crossref PubMed Scopus (1866) Google Scholar, 7Basso E. Fante L. Fowlkes J. Petronilli V. Forte M.A. Bernardi P. J. Biol. Chem. 2005; 280: 18558-18561Abstract Full Text Full Text PDF PubMed Scopus (684) Google Scholar, 8Schinzel A.C. Takeuchi O. Huang Z. Fisher J.K. Zhou Z. Rubens J. Hetz C. Danial N.N. Moskowitz M.A. Korsmeyer S.J. Proc. Natl. Acad. Sci. U. S. A. 2005; 102: 12005-12010Crossref PubMed Scopus (713) Google Scholar, L. Marzo I. Costantini P. Susin S.A. Zamzami N. Petit P.X. Hirsch F. Goulbern M. Poupon M.F. Miccoli L. Xie Z. Reed J.C. Kroemer G. Oncogene. 1999; 18: 2537-2546Crossref PubMed Scopus (189) Google Scholar, 10Tafani M. Karpinich N.O. Hurster K.A. Pastorino J.G. Schneider T. Russo M.A. Farber J.L. J. Biol. Chem. 2002; 277: 10073-10082Abstract Full Text Full Text PDF PubMed Scopus (105) Google Scholar, S. Tsujimoto Y. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 577-582Crossref PubMed Scopus (265) Google Scholar, 12Scorrano L. Ashiya M. Buttle K. Weiler S. Oakes S.A. Mannella C.A. Korsmeyer S.J. Dev. Cell. 2002; 2: 55-67Abstract Full Text Full Text PDF PubMed Scopus (888) Google Scholar, S. Narita M. Tsujimoto Y. Nature. 1999; 399: 483-487Crossref PubMed Scopus (1928) Google Scholar, S. Matsuoka Y. Shinohara Y. Yoneda Y. Tsujimoto Y. J. Cell Biol. 2001; 152: 237-250Crossref PubMed Scopus (332) Google Scholar, I. Brenner C. Zamzami N. Jürgensmeier J.M. Susin S.A. Vieira H.L. Prévost M.C. Xie Z. Matsuyama S. Reed J.C. Kroemer G. Science. 1998; 281: 2027-2031Crossref PubMed Scopus (1058) Google Scholar, S. A. Biochim. Biophys. Acta. 2002; PubMed Scopus Google Scholar, J.G. N. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar, J.D. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar, A. S. PubMed Scopus Google Scholar, Y. N. M. M. M. Biochem. J. PubMed Scopus Google Scholar, 2000; PubMed Google Scholar, S. J. J. Biol. Chem. 1999; Full Text Full Text PDF PubMed Scopus Google Scholar, K. Y. H. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar, J.L. P. V. E. PubMed Scopus Google Scholar, K. Rück A. Brdiczka D. Halestrap A.P. Biochem. J. 1998; PubMed Scopus Google Scholar, M. S. J.M. J. Biochem. 1998; PubMed Scopus Google Scholar, T. K. M.C. D. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar, M. P. Matsuyama S. Cell Biol. PubMed Scopus Google Scholar, M. A. A. G. Biochim. Biophys. Acta. PubMed Scopus Google Scholar, N. V. N. Cell. Biol. PubMed Scopus Google Scholar, N. V. P. K. N.S. N. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar, A.P. C.P. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, J. Nature. PubMed Scopus Google Scholar, A. O. S. J. Cell Biol. 1999; PubMed Scopus Google Scholar, M. Biochem. Biophys. 2000; PubMed Scopus Google Scholar) and the the cyclophilin D of cyclophilin D was by and and and For the the cyclophilin D K. Y. H. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar) was used as a was into Cyclophilin D and were generated by and and and D protein was as K. Y. H. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar). activity was a as a in a to J.L. P. V. E. PubMed Scopus Google Scholar). used for the Bax was by S. hexokinase II was from The hexokinase II was and The was and found to be to the reported The hexokinase II was generated by and The was into cyclophilin D and hexokinase II were into cells of a a Cell with or PPIase-deficient Cyclophilin generated PPIase-deficient and of cyclophilin D to the activity of cyclophilin D is with reports J.D. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google and activity It has been reported that cyclophilin D binds to the ANT in K. Rück A. Brdiczka D. Halestrap A.P. Biochem. J. 1998; PubMed Scopus Google Scholar, M. S. J.M. J. Biochem. 1998; PubMed Scopus Google indicating that cyclophilin D may with the ANT and the opening of the PT pore. in binding of cyclophilin D to the ANT was of which is consistent with J.D. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar). Therefore, we that the cyclophilin D and endogenous cyclophilin D were overexpressed in cancer cells. The glioma cell amounts of endogenous cyclophilin D protein cell not generated the glioma cell overexpressing wild-type or mutant cyclophilin D in that the cyclophilin D were to mitochondria and It was observed that protein of PPIase-deficient were that of wild-type cyclophilin D and that PPIase-deficient cyclophilin D are or to cells. Cyclophilin D PPIase-deficient Cyclophilin D against the effects of cyclophilin D overexpression on apoptosis by has been reported to Bax-dependent cytochrome c release and apoptosis T. K. M.C. D. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar). in overexpression of wild-type cyclophilin D protected the cells from overexpression of the PPIase-deficient the activation cytochrome c and the Bax mitochondrial by that with PPIase-deficient cyclophilin D were to that the anti-apoptotic activity of cyclophilin D is on activity. To clarify the role of Bax in apoptotic cell we the effects of which is from the of M. P. Matsuyama S. Cell Biol. PubMed Scopus Google on the activation by the cells PPIase-deficient cyclophilin D and with or and was in the pro-apoptotic of that Bax plays a role in the cyclophilin to apoptosis. Cyclophilin D of Hexokinase cyclophilin D is a component of the PT pore, we the pore the cyclophilin cells. that overexpression of wild-type cyclophilin D the of mitochondrial-bound hexokinase II, that of mutant cyclophilin D these pore components and and a mitochondrial are not by overexpression of cyclophilin D. that cyclophilin D hexokinase II mitochondrial binding by activity. in CTZ, which has been reported to mitochondrial binding of hexokinase II J.G. N. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google hexokinase II from When the cells were with CTZ, the anti-apoptotic activity of cyclophilin D was that hexokinase II mitochondrial binding is to the anti-apoptotic activity of cyclophilin D. A Hexokinase II from the effects of CsA, a inhibitor of cyclophilin on hexokinase II mitochondrial released hexokinase II from mitochondria as not also a protein inhibitor of not release hexokinase II from mitochondria, the effects of on be from the hexokinase II release the effects of on Bax-induced apoptosis. It has been reported that Bax-induced apoptosis by hexokinase II from mitochondria J.G. N. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar). that CsA, not the activation by of Moreover, the Bax mitochondrial that Bax-induced apoptosis through of hexokinase II mitochondrial of Cyclophilin D Hexokinase II from and to endogenous cyclophilin D is essential to hexokinase II mitochondrial we the effects of RNA on cyclophilin D. cells with a small RNA to target cyclophilin D in cyclophilin D the of endogenous cyclophilin D protein without effects on cyclophilin A or in with siRNA, of endogenous cyclophilin D were and hexokinase II was released from mitochondria into the cytosol. components of the PT pore such as VDAC and ANT were the amounts of hexokinase II protein were may the protein of hexokinase released from mitochondria M. A. A. G. Biochim. Biophys. Acta. PubMed Scopus Google Scholar). The of endogenous cyclophilin D the activation that of endogenous Bax by activation in cyclophilin D cells. that Bax is essential to the of cells to apoptosis by dysfunction of cyclophilin D. that cyclophilin D is a of Bax-mediated apoptosis and is required to mitochondrial binding of hexokinase II. of Cyclophilin D to Hexokinase II the role of the activity of cyclophilin D in the of hexokinase II mitochondrial we endogenous cyclophilin D and cyclophilin D. For of endogenous cyclophilin D, we the that on the of cyclophilin D in endogenous cyclophilin D was cyclophilin D by the were of wild-type cyclophilin D canceled the hexokinase II release by of endogenous cyclophilin D. of the PPIase-deficient cyclophilin D not the cells from hexokinase II clearly show that the activity of endogenous cyclophilin D is essential to the hexokinase II mitochondrial Cyclophilin D for the of Hexokinase hexokinase II was overexpressed in cancer Bax-dependent apoptosis was J.G. N. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar). To clarify the role of cyclophilin D in the anti-apoptotic activity of hexokinase II, we the effects of cyclophilin D dysfunction on the hexokinase II anti-apoptotic activity. in overexpressed hexokinase II was in mitochondria and activation by Bax of cyclophilin D by released hexokinase II from mitochondria Moreover, the anti-apoptotic of hexokinase II overexpression Inactivation of endogenous cyclophilin D by also hexokinase II mitochondrial binding and Bax-mediated apoptosis the that cyclophilin D is essential to the mitochondrial binding and anti-apoptotic activity of hexokinase II. Cyclophilin D has been considered to be essential factor of the PT pore, cyclophilin D null mitochondria are resistant to pore opening (5Nakagawa T. Shimizu S. Watanabe T. Yamaguchi O. Otsu K. Yamagata H. Inohara H. Kubo T. Tsujimoto Y. Nature. 2005; 434: 652-657Crossref PubMed Scopus (1372) Google Scholar, 6Baines C.P. Kaiser R.A. Purcell N.H. Blair N.S. Osinska H. Hambleton M.A. Brunskill E.W. Sayen M.R. Gottlieb R.A. Dorn II, G.W. Robbins J. Molkentin J.D. Nature. 2005; 434: 658-662Crossref PubMed Scopus (1866) Google Scholar, 7Basso E. Fante L. Fowlkes J. Petronilli V. Forte M.A. Bernardi P. J. Biol. Chem. 2005; 280: 18558-18561Abstract Full Text Full Text PDF PubMed Scopus (684) Google Scholar, 8Schinzel A.C. Takeuchi O. Huang Z. Fisher J.K. Zhou Z. Rubens J. Hetz C. Danial N.N. Moskowitz M.A. Korsmeyer S.J. Proc. Natl. Acad. Sci. U. S. A. 2005; 102: 12005-12010Crossref PubMed Scopus (713) Google Scholar). Once the PT pore opens in vitro, mitochondrial outer membrane and cytochrome c release are Based on these in observations, the opening of the PT pore has been regarded as a trigger of apoptosis. However, recent that cyclophilin D and the opening of the PT pore are required to and cell death are not essential to Bax-induced apoptotic cell death (5Nakagawa T. Shimizu S. Watanabe T. Yamaguchi O. Otsu K. Yamagata H. Inohara H. Kubo T. Tsujimoto Y. Nature. 2005; 434: 652-657Crossref PubMed Scopus (1372) Google Scholar, 6Baines C.P. Kaiser R.A. Purcell N.H. Blair N.S. Osinska H. Hambleton M.A. Brunskill E.W. Sayen M.R. Gottlieb R.A. Dorn II, G.W. Robbins J. Molkentin J.D. Nature. 2005; 434: 658-662Crossref PubMed Scopus (1866) Google Scholar). clearly that cyclophilin D is not a trigger of Bax-induced apoptosis. it has been that cyclophilin D is a not a of apoptosis J.D. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar, A. S. PubMed Scopus Google Scholar, Y. N. M. M. M. Biochem. J. PubMed Scopus Google Scholar). cyclophilin D has the role of activity in the anti-apoptotic J.D. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar) has that cyclophilin D suppresses apoptotic cell death by the of a anti-apoptotic mechanism against apoptosis A. S. PubMed Scopus Google Scholar). Thus, the role of activity in the cyclophilin anti-apoptotic To clarify the role of cyclophilin D in apoptosis, we generated a glioma cell overexpressing wild-type or a PPIase-deficient mutant of cyclophilin D of cyclophilin D protected the cells from Bax-mediated apoptosis, that of mutant cyclophilin D Bax mitochondria cytochrome c and apoptotic cell death or apoptosis, Bax to play a role in apoptosis in cells with cyclophilin D and Moreover, of cyclophilin D by cyclosporin A or also the cells against apoptosis and clearly that cyclophilin D is a of apoptosis and that activity is required for this anti-apoptotic also found that overexpression of cyclophilin D the of mitochondrial-bound hexokinase II, overexpression of cyclophilin D mutant and dysfunction of endogenous cyclophilin D these The cyclophilin of hexokinase II mitochondrial binding is on activity detachment of hexokinase II from mitochondria by the anti-apoptotic of cyclophilin D it that hexokinase II plays essential role in the cyclophilin anti-apoptotic Hexokinase II binds to the channel by a protein called VDAC is a primary component of the PT pore complex, and it is also a binding of a pro-apoptotic Hexokinase II appears to the mitochondrial of Bax by the Bax and VDAC J.G. N. J. Biol. Chem. 2002; 277: Full Text Full Text PDF PubMed Scopus Google Scholar). It has been reported that activation of the pathway the mitochondrial binding of hexokinase II and apoptotic cell death N. V. N. Cell. Biol. PubMed Scopus Google Scholar, N. V. P. K. N.S. N. Cell. Full Text Full Text PDF PubMed Scopus Google Scholar). Therefore, the mitochondrial binding of hexokinase II appears to be essential to the anti-apoptotic mechanism of cancer cells. also observed that overexpressed hexokinase II cells from Bax-mediated apoptotic cell death When endogenous cyclophilin D was the anti-apoptotic activity of hexokinase II was that cyclophilin D is required for the hexokinase II-mediated anti-apoptotic propose that cyclophilin D Bax-induced apoptotic cell death through of the mitochondrial binding of hexokinase II by activity. Hexokinase II is to with VDAC in the outer cyclophilin D is in Thus, it appears that cyclophilin D with hexokinase II. Cyclophilin D the hexokinase II mitochondrial binding via interactions with component of the PT pore such as the It has been believed that cyclophilin D pore opening by with the ANT A.P. C.P. J. Biol. Chem. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar). However, a recent J. Nature. PubMed Scopus Google Scholar) this it reports that mitochondrial is observed in mitochondria from The also show that mitochondria are to the ANT the ANT null mitochondria are that the ANT the pore although it is not essential to the opening of the PT pore. with this it was reported that apoptosis is by of cyclophilin D A. S. PubMed Scopus Google Scholar, A. O. S. J. Cell Biol. 1999; PubMed Scopus Google Scholar). Furthermore, the ANT the of hexokinase M. Biochem. Biophys. 2000; PubMed Scopus Google Scholar) and the binding of hexokinase to the in M. L. D. G. J. D. Brdiczka D. Biochim. Biophys. Acta. PubMed Scopus Google Scholar). the cyclophilin D and the ANT be considered factor in pore function. Cyclophilin D to the PT pore and to hexokinase II mitochondrial binding by binding may be the cancer of cyclophilin D has been observed A. S. PubMed Scopus Google Scholar, J.C. Wang J.M. 2002; PubMed Scopus Google Scholar). that cyclophilin D plays role in cancer cells. recent of cyclophilin D that cyclophilin D null mice and show (5Nakagawa T. Shimizu S. Watanabe T. Yamaguchi O. Otsu K. Yamagata H. Inohara H. Kubo T. Tsujimoto Y. Nature. 2005; 434: 652-657Crossref PubMed Scopus (1372) Google Scholar, 6Baines C.P. Kaiser R.A. Purcell N.H. Blair N.S. Osinska H. Hambleton M.A. Brunskill E.W. Sayen M.R. Gottlieb R.A. Dorn II, G.W. Robbins J. Molkentin J.D. Nature. 2005; 434: 658-662Crossref PubMed Scopus (1866) Google Scholar, 7Basso E. Fante L. Fowlkes J. Petronilli V. Forte M.A. Bernardi P. J. Biol. Chem. 2005; 280: 18558-18561Abstract Full Text Full Text PDF PubMed Scopus (684) Google Scholar, 8Schinzel A.C. Takeuchi O. Huang Z. Fisher J.K. Zhou Z. Rubens J. Hetz C. Danial N.N. Moskowitz M.A. Korsmeyer S.J. Proc. Natl. Acad. Sci. U. S. A. 2005; 102: 12005-12010Crossref PubMed Scopus (713) Google Scholar). Moreover, cyclophilin D out not primary and fibroblasts to pro-apoptotic (5Nakagawa T. Shimizu S. Watanabe T. Yamaguchi O. Otsu K. Yamagata H. Inohara H. Kubo T. Tsujimoto Y. Nature. 2005; 434: 652-657Crossref PubMed Scopus (1372) Google Scholar, 6Baines C.P. Kaiser R.A. Purcell N.H. Blair N.S. Osinska H. Hambleton M.A. Brunskill E.W. Sayen M.R. Gottlieb R.A. Dorn II, G.W. Robbins J. Molkentin J.D. Nature. 2005; 434: 658-662Crossref PubMed Scopus (1866) Google Scholar). may be by in the amounts of hexokinase II primary cells and cells. this it has been reported that hexokinase II is highly in it is found in and in amounts S. A. Biochim. Biophys. Acta. 2002; PubMed Scopus Google Scholar). It that cyclophilin D is not essential to cell and plays a pivotal role in the of cancer cells. Therefore, it is that inhibitor of cyclophilin D be not for cells and cyclophilin D may be to as target in However, cyclophilin D inhibitors such as cyclosporin A and sanglifehrin A cellular cyclophilin which cellular inhibitors of cyclophilin D are as S. for N. T. and I. for Y. and for with

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