Functions of neutral ceramidase in the Golgi apparatus
Ceramidases hydrolyze ceramides into sphingosine and fatty acids, with sphingosine being further metabolized into sphingosine-1-phosphate (S1P); thus, ceramidases control the levels of these bioactive sphingolipids in cells and tissues. Neutral ceramidase (nCDase) is highly expressed in colorectal t...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2018-11-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520309123 |
| _version_ | 1818871361588690944 |
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| author | Wataru Sakamoto Nicolas Coant Daniel Canals Lina M. Obeid Yusuf A. Hannun |
| author_facet | Wataru Sakamoto Nicolas Coant Daniel Canals Lina M. Obeid Yusuf A. Hannun |
| author_sort | Wataru Sakamoto |
| collection | DOAJ |
| description | Ceramidases hydrolyze ceramides into sphingosine and fatty acids, with sphingosine being further metabolized into sphingosine-1-phosphate (S1P); thus, ceramidases control the levels of these bioactive sphingolipids in cells and tissues. Neutral ceramidase (nCDase) is highly expressed in colorectal tissues, and a recent report showed that nCDase activity is involved in Wnt/β-catenin signaling. In addition, the inhibition of nCDase decreases the development and progression of colorectal tumor growth. Here, to determine the action of nCDase in colorectal cancer cells, we focused on the subcellular localization and metabolic functions of this enzyme in HCT116 cells. nCDase was found to be located in both the plasma membrane and in the Golgi apparatus, but it had minimal effects on basal levels of ceramide, sphingosine, or S1P. Cells overexpressing nCDase were protected from the cell death and Golgi fragmentation induced by C6-ceramide, and they showed reduced levels of C6-ceramide and higher levels of S1P and sphingosine. Furthermore, compartment-specific metabolic functions of the enzyme were probed using C6-ceramide and Golgi-targeted bacterial SMase (bSMase) and bacterial ceramidase (bCDase). The results showed that Golgi-specific bCDase also demonstrated resistance against the cell death stimulated by C6-ceramide, and it catalyzed the metabolism of ceramides and produced sphingosine in the Golgi. Targeting bSMase to the Golgi resulted in increased levels of ceramide that were attenuated by the expression of nCDase, also supporting its ability to metabolize Golgi-generated ceramide. These results are critical in understanding the functions of nCDase actions in colorectal cancer cells as well as the compartmentalized pathways of sphingolipid metabolism. |
| first_indexed | 2024-12-19T12:21:42Z |
| format | Article |
| id | doaj.art-925efbb571be4691aa94e30b82fee652 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-19T12:21:42Z |
| publishDate | 2018-11-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-925efbb571be4691aa94e30b82fee6522022-12-21T20:21:44ZengElsevierJournal of Lipid Research0022-22752018-11-01591121162125Functions of neutral ceramidase in the Golgi apparatusWataru Sakamoto0Nicolas Coant1Daniel Canals2Lina M. Obeid3Yusuf A. Hannun4Department of Medicine, Stony Brook University, Stony Brook, NY; Stony Brook Cancer Center, Stony Brook University, Stony Brook, NY; Exploratory Research Laboratories, Ono Pharmaceutical Co., Ltd., Osaka, JapanDepartment of Medicine, Stony Brook University, Stony Brook, NY; Stony Brook Cancer Center, Stony Brook University, Stony Brook, NYDepartment of Medicine, Stony Brook University, Stony Brook, NY; Stony Brook Cancer Center, Stony Brook University, Stony Brook, NYDepartment of Medicine, Stony Brook University, Stony Brook, NY; Stony Brook Cancer Center, Stony Brook University, Stony Brook, NY; Northport Veterans Affairs Medical Center, Northport, NYTo whom correspondence should be addressed.; Department of Medicine, Stony Brook University, Stony Brook, NY; Stony Brook Cancer Center, Stony Brook University, Stony Brook, NY; Department of Biochemistry, Stony Brook University, Stony Brook, NY; Department of Pharmacology, Stony Brook University, Stony Brook, NY; Department of Pathology, Stony Brook University, Stony Brook, NYCeramidases hydrolyze ceramides into sphingosine and fatty acids, with sphingosine being further metabolized into sphingosine-1-phosphate (S1P); thus, ceramidases control the levels of these bioactive sphingolipids in cells and tissues. Neutral ceramidase (nCDase) is highly expressed in colorectal tissues, and a recent report showed that nCDase activity is involved in Wnt/β-catenin signaling. In addition, the inhibition of nCDase decreases the development and progression of colorectal tumor growth. Here, to determine the action of nCDase in colorectal cancer cells, we focused on the subcellular localization and metabolic functions of this enzyme in HCT116 cells. nCDase was found to be located in both the plasma membrane and in the Golgi apparatus, but it had minimal effects on basal levels of ceramide, sphingosine, or S1P. Cells overexpressing nCDase were protected from the cell death and Golgi fragmentation induced by C6-ceramide, and they showed reduced levels of C6-ceramide and higher levels of S1P and sphingosine. Furthermore, compartment-specific metabolic functions of the enzyme were probed using C6-ceramide and Golgi-targeted bacterial SMase (bSMase) and bacterial ceramidase (bCDase). The results showed that Golgi-specific bCDase also demonstrated resistance against the cell death stimulated by C6-ceramide, and it catalyzed the metabolism of ceramides and produced sphingosine in the Golgi. Targeting bSMase to the Golgi resulted in increased levels of ceramide that were attenuated by the expression of nCDase, also supporting its ability to metabolize Golgi-generated ceramide. These results are critical in understanding the functions of nCDase actions in colorectal cancer cells as well as the compartmentalized pathways of sphingolipid metabolism.http://www.sciencedirect.com/science/article/pii/S0022227520309123neutral ceramidasesphingolipidsGolgiapoptosiscolon cancer |
| spellingShingle | Wataru Sakamoto Nicolas Coant Daniel Canals Lina M. Obeid Yusuf A. Hannun Functions of neutral ceramidase in the Golgi apparatus Journal of Lipid Research neutral ceramidase sphingolipids Golgi apoptosis colon cancer |
| title | Functions of neutral ceramidase in the Golgi apparatus |
| title_full | Functions of neutral ceramidase in the Golgi apparatus |
| title_fullStr | Functions of neutral ceramidase in the Golgi apparatus |
| title_full_unstemmed | Functions of neutral ceramidase in the Golgi apparatus |
| title_short | Functions of neutral ceramidase in the Golgi apparatus |
| title_sort | functions of neutral ceramidase in the golgi apparatus |
| topic | neutral ceramidase sphingolipids Golgi apoptosis colon cancer |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520309123 |
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