Analysis of lipid-composition changes in plasma membrane microdomains[S]
Sphingolipids accumulate in plasma membrane microdomain sites, such as caveolae or lipid rafts. Such microdomains are considered to be important nexuses for signal transduction, although changes in the microdomain lipid components brought about by signaling are poorly understood. Here, we applied a...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2015-08-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S002222752035536X |
| _version_ | 1818735004743630848 |
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| author | Hideo Ogiso Makoto Taniguchi Toshiro Okazaki |
| author_facet | Hideo Ogiso Makoto Taniguchi Toshiro Okazaki |
| author_sort | Hideo Ogiso |
| collection | DOAJ |
| description | Sphingolipids accumulate in plasma membrane microdomain sites, such as caveolae or lipid rafts. Such microdomains are considered to be important nexuses for signal transduction, although changes in the microdomain lipid components brought about by signaling are poorly understood. Here, we applied a cationic colloidal silica bead method to analyze plasma membrane lipids from monolayer cells cultured in a 10 cm dish. The detergent-resistant fraction from the silica bead-coated membrane was analyzed by LC-MS/MS to evaluate the microdomain lipids. This method revealed that glycosphingolipids composed the microdomains as a substitute for sphingomyelin (SM) in mouse embryonic fibroblasts (tMEFs) from an SM synthase 1/2 double KO (DKO) mouse. The rate of formation of the detergent-resistant region was unchanged compared with that of WT-tMEFs. C2-ceramide (Cer) stimulation caused greater elevations in diacylglycerol and phosphatidic acid levels than in Cer levels within the microdomains of WT-tMEFs. We also found that lipid changes in the microdomains of SM-deficient DKO-tMEFs caused by serum stimulation occurred in the same manner as that of WT-tMEFs. This practical method for analyzing membrane lipids will facilitate future comprehensive analyses of membrane microdomain-associated responses. |
| first_indexed | 2024-12-18T00:14:22Z |
| format | Article |
| id | doaj.art-c2d659a6c7504e3c8a8010451d2a92c4 |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-18T00:14:22Z |
| publishDate | 2015-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-c2d659a6c7504e3c8a8010451d2a92c42022-12-21T21:27:33ZengElsevierJournal of Lipid Research0022-22752015-08-0156815941605Analysis of lipid-composition changes in plasma membrane microdomains[S]Hideo Ogiso0Makoto Taniguchi1Toshiro Okazaki2Department of Hematology/Immunology Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293, JapanMedical Research Institute, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293, JapanTo whom correspondence should be addressed; Department of Hematology/Immunology Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293, Japan; Medical Research Institute, Kanazawa Medical University, 1-1 Daigaku, Uchinada, Ishikawa 920-0293, JapanSphingolipids accumulate in plasma membrane microdomain sites, such as caveolae or lipid rafts. Such microdomains are considered to be important nexuses for signal transduction, although changes in the microdomain lipid components brought about by signaling are poorly understood. Here, we applied a cationic colloidal silica bead method to analyze plasma membrane lipids from monolayer cells cultured in a 10 cm dish. The detergent-resistant fraction from the silica bead-coated membrane was analyzed by LC-MS/MS to evaluate the microdomain lipids. This method revealed that glycosphingolipids composed the microdomains as a substitute for sphingomyelin (SM) in mouse embryonic fibroblasts (tMEFs) from an SM synthase 1/2 double KO (DKO) mouse. The rate of formation of the detergent-resistant region was unchanged compared with that of WT-tMEFs. C2-ceramide (Cer) stimulation caused greater elevations in diacylglycerol and phosphatidic acid levels than in Cer levels within the microdomains of WT-tMEFs. We also found that lipid changes in the microdomains of SM-deficient DKO-tMEFs caused by serum stimulation occurred in the same manner as that of WT-tMEFs. This practical method for analyzing membrane lipids will facilitate future comprehensive analyses of membrane microdomain-associated responses.http://www.sciencedirect.com/science/article/pii/S002222752035536Xcationic colloidal silica beadsliquid chromatography-tandem mass spectrometrylipid raftlipidomicsdiacylglycerolphosphatidic acid |
| spellingShingle | Hideo Ogiso Makoto Taniguchi Toshiro Okazaki Analysis of lipid-composition changes in plasma membrane microdomains[S] Journal of Lipid Research cationic colloidal silica beads liquid chromatography-tandem mass spectrometry lipid raft lipidomics diacylglycerol phosphatidic acid |
| title | Analysis of lipid-composition changes in plasma membrane microdomains[S] |
| title_full | Analysis of lipid-composition changes in plasma membrane microdomains[S] |
| title_fullStr | Analysis of lipid-composition changes in plasma membrane microdomains[S] |
| title_full_unstemmed | Analysis of lipid-composition changes in plasma membrane microdomains[S] |
| title_short | Analysis of lipid-composition changes in plasma membrane microdomains[S] |
| title_sort | analysis of lipid composition changes in plasma membrane microdomains s |
| topic | cationic colloidal silica beads liquid chromatography-tandem mass spectrometry lipid raft lipidomics diacylglycerol phosphatidic acid |
| url | http://www.sciencedirect.com/science/article/pii/S002222752035536X |
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