Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS)
Lipids, such as phosphoinositides (PIPs) and diacylglycerol (DAG), are important signaling intermediates involved in cellular processes such as T cell receptor (TCR)-mediated signal transduction. Here we report identification and quantification of PIP, PIP2 and DAG from crude lipid extracts. Capital...
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| Format: | Article |
| Language: | English |
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MDPI AG
2012-01-01
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| Series: | Metabolites |
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| Online Access: | http://www.mdpi.com/2218-1989/2/1/57 |
| _version_ | 1818190775976984576 |
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| author | Mathias Haag Angelika Schmidt Timo Sachsenheimer Britta Brügger |
| author_facet | Mathias Haag Angelika Schmidt Timo Sachsenheimer Britta Brügger |
| author_sort | Mathias Haag |
| collection | DOAJ |
| description | Lipids, such as phosphoinositides (PIPs) and diacylglycerol (DAG), are important signaling intermediates involved in cellular processes such as T cell receptor (TCR)-mediated signal transduction. Here we report identification and quantification of PIP, PIP2 and DAG from crude lipid extracts. Capitalizing on the different extraction properties of PIPs and DAGs allowed us to efficiently recover both lipid classes from one sample. Rapid analysis of endogenous signaling molecules was performed by nano-electrospray ionization tandem mass spectrometry (nano-ESI MS/MS), employing lipid class-specific neutral loss and multiple precursor ion scanning for their identification and quantification. Profiling of DAG, PIP and PIP2 molecular species in primary human T cells before and after TCR stimulation resulted in a two-fold increase in DAG levels with a shift towards 1-stearoyl-2-arachidonoyl-DAG in stimulated cells. PIP2 levels were slightly reduced, while PIP levels remained unchanged. |
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| id | doaj.art-7be6cfe6a28f40ab94fa6067a6adc673 |
| institution | Directory Open Access Journal |
| issn | 2218-1989 |
| language | English |
| last_indexed | 2024-12-12T00:04:05Z |
| publishDate | 2012-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Metabolites |
| spelling | doaj.art-7be6cfe6a28f40ab94fa6067a6adc6732022-12-22T00:45:09ZengMDPI AGMetabolites2218-19892012-01-0121577610.3390/metabo2010057Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS)Mathias HaagAngelika SchmidtTimo SachsenheimerBritta BrüggerLipids, such as phosphoinositides (PIPs) and diacylglycerol (DAG), are important signaling intermediates involved in cellular processes such as T cell receptor (TCR)-mediated signal transduction. Here we report identification and quantification of PIP, PIP2 and DAG from crude lipid extracts. Capitalizing on the different extraction properties of PIPs and DAGs allowed us to efficiently recover both lipid classes from one sample. Rapid analysis of endogenous signaling molecules was performed by nano-electrospray ionization tandem mass spectrometry (nano-ESI MS/MS), employing lipid class-specific neutral loss and multiple precursor ion scanning for their identification and quantification. Profiling of DAG, PIP and PIP2 molecular species in primary human T cells before and after TCR stimulation resulted in a two-fold increase in DAG levels with a shift towards 1-stearoyl-2-arachidonoyl-DAG in stimulated cells. PIP2 levels were slightly reduced, while PIP levels remained unchanged.http://www.mdpi.com/2218-1989/2/1/57T cell signalingsignaling lipidsdiacylglycerolphosphoinositideslipidomicsnano-electrospray ionization tandem mass spectrometry |
| spellingShingle | Mathias Haag Angelika Schmidt Timo Sachsenheimer Britta Brügger Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS) Metabolites T cell signaling signaling lipids diacylglycerol phosphoinositides lipidomics nano-electrospray ionization tandem mass spectrometry |
| title | Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS) |
| title_full | Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS) |
| title_fullStr | Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS) |
| title_full_unstemmed | Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS) |
| title_short | Quantification of Signaling Lipids by Nano-Electrospray Ionization Tandem Mass Spectrometry (Nano-ESI MS/MS) |
| title_sort | quantification of signaling lipids by nano electrospray ionization tandem mass spectrometry nano esi ms ms |
| topic | T cell signaling signaling lipids diacylglycerol phosphoinositides lipidomics nano-electrospray ionization tandem mass spectrometry |
| url | http://www.mdpi.com/2218-1989/2/1/57 |
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