PMID- 21613224 OWN - NLM STAT- MEDLINE DCOM- 20120124 LR - 20211020 IS - 1083-351X (Electronic) IS - 0021-9258 (Print) IS - 0021-9258 (Linking) VI - 286 IP - 28 DP - 2011 Jul 15 TI - Novel pathway of ceramide production in mitochondria: thioesterase and neutral ceramidase produce ceramide from sphingosine and acyl-CoA. PG - 25352-62 LID - 10.1074/jbc.M110.214866 [doi] AB - Reports suggest that excessive ceramide accumulation in mitochondria is required to initiate the intrinsic apoptotic pathway and subsequent cell death, but how ceramide accumulates is unclear. Here we report that liver mitochondria exhibit ceramide formation from sphingosine and palmitoyl-CoA and from sphingosine and palmitate. Importantly, this activity was markedly decreased in liver from neutral ceramidase (NCDase)-deficient mice. Moreover, the levels of ceramide were dissimilar in liver mitochondria of WT and NCDase KO mice. These results suggest that NCDase is a key participant of ceramide formation in liver mitochondria. We also report that highly purified liver mitochondria have ceramidase, reverse ceramidase, and thioesterase activities. Increased accessibility of palmitoyl-CoA to the mitochondrial matrix with the pore-forming peptide zervamicin IIB resulted in 2-fold increases in palmitoyl-CoA hydrolysis by thioesterase. This increased hydrolysis was accompanied by an increase in ceramide formation, demonstrating that both outer membrane and matrix localized thioesterases can regulate ceramide formation. Also, ceramide formation might occur both in the outer mitochondrial membrane and in the mitochondrial matrix, suggesting the existence of distinct ceramide pools. Taken together, these results suggest that the reverse activity of NCDase contributes to sphingolipid homeostasis in this organelle in vivo. FAU - Novgorodov, Sergei A AU - Novgorodov SA AD - Ralph H. Johnson Veterans Affairs Medical Center, Charleston, South Carolina 29401, USA. FAU - Wu, Bill X AU - Wu BX FAU - Gudz, Tatyana I AU - Gudz TI FAU - Bielawski, Jacek AU - Bielawski J FAU - Ovchinnikova, Tatiana V AU - Ovchinnikova TV FAU - Hannun, Yusuf A AU - Hannun YA FAU - Obeid, Lina M AU - Obeid LM LA - eng GR - R01 CA087584/CA/NCI NIH HHS/United States GR - AG16583/AG/NIA NIH HHS/United States GR - C06 RR018823/RR/NCRR NIH HHS/United States GR - UL1 RR029882/RR/NCRR NIH HHS/United States GR - P20RR17677-04/RR/NCRR NIH HHS/United States GR - P20 RR017677/RR/NCRR NIH HHS/United States GR - CA87584/CA/NCI NIH HHS/United States GR - R01 AG016583/AG/NIA NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't PT - Research Support, U.S. Gov't, Non-P.H.S. DEP - 20110525 PL - United States TA - J Biol Chem JT - The Journal of biological chemistry JID - 2985121R RN - 0 (Ceramides) RN - 0 (Mitochondrial Proteins) RN - 0 (Peptaibols) RN - 1763-10-6 (Palmitoyl Coenzyme A) RN - 79395-85-0 (zervamicin IIB) RN - EC 3.1.2.2 (Palmitoyl-CoA Hydrolase) RN - EC 3.5.1.23 (Neutral Ceramidase) RN - NGZ37HRE42 (Sphingosine) SB - IM MH - Animals MH - Ceramides/genetics/*metabolism MH - Hydrolysis/drug effects MH - Lipid Metabolism/drug effects/*physiology MH - Male MH - Mice MH - Mice, Knockout MH - Mitochondria, Liver/*enzymology/genetics MH - Mitochondrial Proteins/genetics/*metabolism MH - Neutral Ceramidase/genetics/*metabolism MH - Palmitoyl Coenzyme A/genetics/*metabolism MH - Palmitoyl-CoA Hydrolase MH - Peptaibols/pharmacology MH - Rats MH - Rats, Sprague-Dawley MH - Sphingosine/genetics/*metabolism PMC - PMC3137106 EDAT- 2011/05/27 06:00 MHDA- 2012/01/25 06:00 PMCR- 2012/07/15 CRDT- 2011/05/27 06:00 PHST- 2011/05/27 06:00 [entrez] PHST- 2011/05/27 06:00 [pubmed] PHST- 2012/01/25 06:00 [medline] PHST- 2012/07/15 00:00 [pmc-release] AID - S0021-9258(19)48694-8 [pii] AID - M110.214866 [pii] AID - 10.1074/jbc.M110.214866 [doi] PST - ppublish SO - J Biol Chem. 2011 Jul 15;286(28):25352-62. doi: 10.1074/jbc.M110.214866. Epub 2011 May 25.