Abstract
Among the permeability pathways in the mitochondrial outer membrane (MOM), whose elucidation was pioneered by Kathleen Kinnally, there is one formed by the lipid, ceramide. Electron microscopic visualization shows that ceramide channels are large cylindrical structures of varying pore size, with a most frequent size of 10 nm in diameter, large enough to allow all soluble proteins to translocate between the cytosol and the mitochondrial intermembrane space. Similar results were obtained with electrophysiological measurements. Studies of the dynamics of the channels are consistent with a right cylinder. Ceramide channels form at mole fractions of ceramide that are found in the MOM early in the apoptotic process, before or at the time of protein release from mitochondria. That these channels are good candidates for the protein release pathway is supported by the fact that channel formation is inhibited by anti-apoptotic proteins and favored by Bax. Bcl-xL inhibits ceramide channel formation by binding to the apolar ceramide tails using its hydrophobic grove. Bax interaction with the polar regions of ceramide results in MOM permeabilization through synergy with ceramide. Evidence that ceramide channels actually function to favor apoptosis in vivo is supported by the expression of Bcl-xL containing point mutations in cells induced to undergo apoptosis. The Bcl-xL mutants inhibit differentially Bax and ceramide channels and thus tease apart, to some extent, these two modes of MOM permeabilization. Ceramide channels have the right properties and appropriate regulation to be key players in the induction of apoptosis.
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References
Andrews NW, Almeida PE, Corrotte M (2014) Damage control: cellular mechanisms of plasma membrane repair. Trends Cell Biol 24:734–742
Anishkin A, Sukharev S, Colombini M (2006) Searching for the molecular arrangement of transmembrane ceramide channels. Biophys J 90:2414–2426
Belaud-Rotureau MA, Leducq N, Macouillard Poulletier de Gannes F, Diolez P, Lacoste L, Lacombe F, Bernard P, Belloc F (2000) Early transitory rise in intracellular pH leads to Bax conformation change during ceramide-induced apoptosis. Apoptosis 5:551–560
Beverly LJ, Howell LA, Hernandez-Corbacho M, Casson L, Chipuk JE, Siskind LJ (2013) BAK activation is necessary and sufficient to drive ceramide synthase-dependent ceramide accumulation following inhibition of BCL2-like proteins. Biochem J 452(1):111–119
Birbes H, El Bawab S, Hannun YA, et al. (2001) Selective hydrolysis of a mitochondrial pool of sphingomyelin induces apoptosis. FASEB J 15:2669–2679
Birbes H, Luberto C, Hsu YT, et al. (2005) A mitochondrial pool of sphingomyelin is involved in TNFalpha-induced Bax translocation to mitochondria. Biochem J 386:445–451
Castro BM, Prieto M, Silva LC (2014) Ceramide: a simple sphingolipid with unique biophysical properties. Prog Lipid Res 54:53–67
Chang KT, Anishkin A, Patwardhan GA, Beverly LJ, Siskind LJ, Colombini M (2015) Ceramide channels: destabilization by Bcl-xL and role in apoptosis. Biochim Biophys Acta 1848:2374–2384
Colombini M (1979) Candidate for the permeability pathway of the outer mitochondrial-membrane. Nature 279:643–645
Colombini M (2013) Membrane channels formed by ceramide. Handb Exp Pharmacol 215:109–126
Dejean LM, Ryu SY, Martinez-Caballero S, Teijido O, Peixoto PM, Kinnally KW (2010) MAC and Bcl-2 family proteins conspire in a deadly plot. Biochim Biophys Acta 1797:1231–1238
Elrick MJ, Fluss S, Colombini M (2006) Sphingosine, a product of ceramide hydrolysis by ceramidase, disassembles ceramide channels. Biophys J 91:1749–1756
Ganesan V, Perera MN, Colombini D, Datskovskiy D, Chadha K, Colombini M (2010) Ceramide and activated Bax act synergistically to permeabilize the mitochondrial outer membrane. Apoptosis 15:553–562
Gillies LA, Kuwana T (2014) Apoptosis regulation at the mitochondrial outer membrane. J Cell Biochem 115:632–640
Green DR, Galluzzi L, Kroemer G (2014) Metabolic control of cell death. Science 345(6203):1250256
Hannun YA, Obeid LM (2008) Principles of bioactive lipid signaling: lessons from sphingolipids. Nat. Rev Mol Cell Biol 9:139–150
Hannun YA, Obeid LM (2011) Many ceramides. J Biol Chem 286:27855–27862
Jin J, Hou Q, Mullen TD, Zeidan YH, Bielawski J, Kraveka JM, Bielawska A, Obeid LM, Hannun YA, Hsu YT (2008) Ceramide generated by sphingomyelin hydrolysis and the salvage pathway is involved in hypoxia/reoxygenation-induced Bax redistribution to mitochondria in NT-2 cells. J Biol Chem 283:26509–26517
Khavandgar Z, Murshed M (2015) Sphingolipid metabolism and its role in the skeletal tissues. Cell Mol Life Sci 72:959–969
Kim HJ, Mun JY, Chun YJ, Choi KH, Kim MY (2001) Bax-dependent apoptosis induced by ceramide in HL-60 cells. FEBS Lett 505:264–268
Kinnally KW, Muro C, Campo ML (2000) MCC and PSC, the putative protein import channels of mitochondria. J Bioenerg Biomembr 32:47–54
Kinnally KW, Antonsson B (2007) A tale of two mitochondrial channels, MAC and PTP, in apoptosis. Apoptosis 12:857–868
Kogot-Levin A, Saada A (2014) Ceramide and the mitochondrial respiratory chain. Biochimie 100:88–94
Kroesen BJ, Jacobs S, Pettus BJ, Sietsma H, Kok JW, YA H, de Leij L F (2003) BcR-induced apoptosis involves differential regulation of C16 and C24-ceramide formation and sphingolipid-dependent activation of the proteasome. J Biol Chem 278:14723–14731
Kroesen BJ, Pettus B, Luberto C, Busman M, Sietsma H, de Leij L, Hannun YA (2001) Induction of apoptosis through B-cell receptor cross-linking occurs via de novo generated C16-ceramide and involves mitochondria. J Biol Chem 276:13606–13614
Lee EF, Czabotar PE, Smith BJ, Deshayes K, Zobel K, Colman PM, Fairlie WD (2007) Crystal structure of abt-737 complexed with bcl-xl: implications for selectivity of antagonists of the bcl-2 family. Cell Death Differ 14:1711–1713
Lee H, Rotolo JA, Mesicek J, Penate-Medina T, Rimner A, Liao WC, Yin X, Ragupathi G, Ehleiter D, Gulbins E, Zhai D, Reed JC, Haimovitz-Friedman A, Fuks Z, Kolesnick R (2011) Mitochondrial ceramide-rich macrodomains functionalize Bax upon irradiation. PLoS One 6(6):e19783
Martínez-Abundis E, Correa F, Pavón N, Zazueta C (2009) Bax distribution into mitochondrial detergent-resistant microdomains is related to ceramide and cholesterol content in postischemic hearts. Febs J 276:5579–5588
Meckfessel MH, Brandt S (2014) The structure, function, and importance of ceramides in skin and their use as therapeutic agents in skin-care products. J Am Acad Dermatol 71:177–184
Mesicek J, Lee H, Feldman T, Jiang X, Skobeleva A, Berdyshev EV, Haimovitz-Friedman A, Fuks Z, Kolesnick R (2010) Ceramide synthases 2, 5, and 6 confer distinct roles in radiation-induced apoptosis in HeLa cells. Cell Signal 22:1300–1307
Moldoveanu T, Follis AV, Kriwacki RW, Green DR (2014) Many players in BCL-2 family affairs. Trends Biochem Sci 39:101–111
Nikaido H, Song SA, Shaltiel L, Nurminen M (1976) Outer membrane of salmonella XIV. Reduced transmembrane diffusion rates in porin-deficient mutants. Biochem Biophys Res Commun 76:324–330
Obeid LM, Linardic CM, Karolak LA, Hannun YA (1993) Programmed cell death induced by ceramide. Science 259:1769–1771
Pastorino JG, Tafani M, Rothman RJ, Marcinkeviciute A, Hoek JB, Farber JL (1999) Functional consequences of the sustained or transient activation by Bax of the mitochondrial permeability transition pore. J Biol Chem 274:31734–31739
Patwardhan GA, Beverly LJ, Siskind LJ (2015) Sphingolipids and mitochondrial apoptosis. J Bioenerg Biomembr. doi:10.1007/s10863-015-9602-3
Peixoto PM, Dejean LM, Kinnally KW (2012) The therapeutic potential of mitochondrial channels in cancer, ischemia-reperfusion injury, and neurodegeneration. Mitochondrion 12:14–23
Perera MN, Ganesan V, Siskind LJ, Szulc ZM, Bielawski J, Bielawska A, Bittman R, Colombini M (2012a) Ceramide channels: influence of molecular structure on channel formation in membranes. Biochim Biophys Acta 1818:1291–1301
Perera MN, Ganesan V, Siskind LJ, Szulc ZM, Bielawska A, Bittman R, Colombini, M (2016) Ceramide channel: structural basis for selective membrane targeting. Chem Phys Lipids 194:110–116
Perera MN, Lin SH, Peterson YK, Bielawska A, Szulc ZM, Bittman R, Colombini M (2012b) BAX, Bcl-xL exert their regulation on different sites of the ceramide channel. Biochem J 445:81–91
Rasola A, Bernardi P (2011) Mitochondrial permeability transition in Ca(2+)-dependent apoptosis and necrosis. Cell Calcium 50:222–233
Renault TT, Manon S (2011) Bax: addressed to kill. Biochimie 93:1379–1391
Rodriguez-Lafrasse C, Alphonse G, Broquet P, Aloy MT, Louisot P, Rousson R (2001) Temporal relationships between ceramide production, caspase activation and mitochondrial dysfunction in cell lines with varying sensitivity to anti-Fas-induced apoptosis. Biochem J 357(Pt 2):407–416
Rotolo JA, Zhang J, Donepudi M, Lee H, Fuks Z, Kolesnick R (2005) Caspase-dependent and -independent activation of acid sphingomyelinase signaling. J Biol Chem 280:26425–26434
Samanta S, Stiban J, Maugel TK, et al. (2011) Visualization of ceramide channels by transmission electron microscopy. Biochim Biophys Acta 1808:1196–1201
Schein SJ, Colombini M, Finkelstein A (1976) Reconstitution in planar lipid bilayers of a voltage-dependent anion-selective channel obtained from paramecium mitochondria. J Membr Biol 30:99–120
Shao C, Sun B, Colombini M, DeVoe D (2012) Dynamics of ceramide channels detected using a microfluidic system. PLoS One 7(9):e43513
Siskind LJ, Colombini M (2000) The lipids C2- and C16-ceramide form large stable channels: implications for apoptosis. J Biol Chem 275:38640–38644
Siskind LJ, Davoody A, Lewin N, et al. (2003) Enlargement and contracture of C2-ceramide channels. Biophys J 85:1560–1575
Siskind LJ, Feinstein L, Yu T, Davis JS, Jones D, Choi J, Zuckerman JE, Tan W, Hill RB, Hardwick JM, Colombini M (2008) Anti-apoptotic Bcl-2 family proteins disassemble ceramide channels. J Biol Chem 283:6622–6630
Siskind LJ, Fluss S, Bui M, Colombini M (2005) Sphingosine forms channels in membranes that differ greatly from those formed by ceramide. J Bioenerg Biomembr 37:227–236
Siskind LJ, Kolesnick RN, Colombini M (2002) Ceramide channels increase the permeability of the mitochondrial outer membrane to small proteins. J Biol Chem 277:26796–26803
Siskind LJ, Kolesnick RN, Colombini M (2006) Ceramide forms channels in mitochondrial outer membranes at physiologically relevant concentrations. Mitochondrion 6:118–125
Siskind LJ, Mullen TD, Rosales KR, Clarke CJ, Hernandez-Corbacho MJ, Edinger AL, Obeid LM (2010) The Bcl-2 protein BAK is required for long-chain ceramide generation during apoptosis. J Biol Chem 285:11818–11826
Stiban J, Fistere Jr D, Colombini M (2006) Dihydroceramide hinders ceramide channel formation: implications on apoptosis. Apoptosis 11:773–780
Stiban J, Perera M (2015) Very long chain ceramides interfere with C16-ceramide-induced channel formation: a plausible mechanism for regulating the initiation of intrinsic apoptosis. Biochim Biophys Acta 1848:561–567
Tedeschi H, Kinnally KW (1987) Channels in the mitochondrial outer-membrane - evidence from patch clamp studies. J Bioenerg Biomembr 19:321–327
Thomas Jr RL, Matsko CM, Lotze MT, Amoscato AA (1999) Mass spectrometric identification of increased C16 ceramide levels during apoptosis. J Biol Chem 274:30580–30588
von Haefen C, Wieder T, Gillissen B, Stärck L, Graupner V, Dörken B, PT D (2002) Ceramide induces mitochondrial activation and apoptosis via a Bax-dependent pathway in human carcinoma cells. Oncogene 21:4009–4019
Westphal D, Kluck RM, Dewson G (2014) Building blocks of the apoptotic pore: how Bax and Bak are activated and oligomerize during apoptosis. Cell Death Differ 21:196–205
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This work was supported by funding from the National Science Foundation (MCB-1023008).
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Supplemental Figure 1
Panel A. The working model of a ceramide channel consisting of 48 columns of ceramide arranged in an anti-parallel fashion. The phospholipid bilayer surrounding the channel is not illustrated. This forms an aqueous pore that is 10 nm in diameter, the size most commonly observed experimentally. The pore is lined by the polar ends of the ceramide molecules with the oxygens in red. The crystal structure of Bcl-xL is superimposed on one portion of the channel at the bottom of the figure. It is positioned to overlap the apolar tails of a ceramide at the end of one of the columns. Panel B. A pair of ceramide columns in an anti-parallel orientation illustrates the channel structure in detail. Each column consists of 6 ceramide molecules connected by hydrogen bonding (yellow) both through the amide linkages and the twin hydroxyls. Some water molecules are also involved in the hydrogen-bonded network. (Reprinted with permission from Chang et al., 2015) (GIF 205 kb)
Supplemental Figure 2
Metabolic pathways that generate or consume ceramide. (GIF 10 kb)
Supplemental Figure 3
Model of the Bcl-xL/ceramide interaction from in-silico docking experiments. Structure of C16-ceramide, in green, bound to Bcl-xL, shown in ribbon mode. Panel A some of the amino acid side chains: F97, F105 and F146 are shown in full space filling structure and labeled as “F″ so as to illustrate their interaction with the acyl chain of ceramide. Panel B illustrates the polar interactions between the ceramide hydroxyls (shown in space filling mode) and two side chains if Bcl-xL: R100 and Y195. R100 has 2 hydrogen bonds with the oxygen of the hydroxyl on carbon 3 of ceramide (labeled as 3-OH). The hydroxyl of Y195 (labeled as “Y″) is aligned next to the C1 hydroxyl (labeled as 1-OH) of ceramide in such a way that their dipoles are antiparallel. (Reprinted with permission from Chang et al., 2015) (GIF 132 kb)
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Colombini, M. Ceramide channels and mitochondrial outer membrane permeability. J Bioenerg Biomembr 49, 57–64 (2017). https://doi.org/10.1007/s10863-016-9646-z
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DOI: https://doi.org/10.1007/s10863-016-9646-z