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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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Bibliographic Details
Main Authors: Haag, Mathias (Author) , Sachsenheimer, Timo (Author) , Brügger, Britta (Author)
Format: Article (Journal)
Language:English
Published: 16 January 2012
In: Metabolites
Year: 2012, Volume: 2, Issue: 1, Pages: 57-76
ISSN:2218-1989
DOI:10.3390/metabo2010057
Online Access:Verlag, kostenfrei, Volltext: http://dx.doi.org/10.3390/metabo2010057
Verlag, kostenfrei, Volltext: http://www.mdpi.com/2218-1989/2/1/57
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Author Notes:Mathias Haag, Angelika Schmidt, Timo Sachsenheimer, Britta Brügger
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Summary: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.
Item Description:Gesehen am 24.08.2018
Physical Description:Online Resource
ISSN:2218-1989
DOI:10.3390/metabo2010057

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