Fatty-acyl chain profiles of cellular phosphoinositides

Phosphoinositides are rapidly turning-over phospholipids that play key roles in intracellular signaling and modulation of membrane effectors. Through technical refinements we have improved sensitivity in the analysis of the phosphoinositide PI, PIP, and PIP2 pools from living cells using mass spectr...

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Vydáno v:	Biochimica et biophysica acta. Molecular and cell biology of lipids Ročník 1862; číslo 5; s. 513 - 522
Hlavní autoři:	Traynor-Kaplan, Alexis, Kruse, Martin, Dickson, Eamonn J., Dai, Gucan, Vivas, Oscar, Yu, Haijie, Whittington, Dale, Hille, Bertil
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	Netherlands Elsevier B.V 01.05.2017
Témata:	animal ovaries Animals Arachidonic acid butanol cell culture Chinese hamsters CHO Cells Cricetinae Cricetulus derivatization detection limit electrospray ionization mass spectrometry fatty acids Fatty Acids - chemistry Fatty Acids - isolation & purification ganglia Humans kidney cells Lipidomics Mass spectrometry monitoring Phosphatidylinositols - chemistry Phosphatidylinositols - isolation & purification phospholipase C Phospholipids Phospholipids - chemistry Phospholipids - isolation & purification pineal body receptors Signal Transduction sodium Spectrometry, Mass, Electrospray Ionization - methods PLC MRM Lipidomics Phospholipids DAG Arachidonic acid IP3 PIP SCG PI D-PI TQ Mass spectrometry PIP2
ISSN:	1388-1981, 1879-2618
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Popis
Shrnutí:	Phosphoinositides are rapidly turning-over phospholipids that play key roles in intracellular signaling and modulation of membrane effectors. Through technical refinements we have improved sensitivity in the analysis of the phosphoinositide PI, PIP, and PIP2 pools from living cells using mass spectrometry. This has permitted further resolution in phosphoinositide lipidomics from cell cultures and small samples of tissue. The technique includes butanol extraction, derivatization of the lipids, post-column infusion of sodium to stabilize formation of sodiated adducts, and electrospray ionization mass spectrometry in multiple reaction monitoring mode, achieving a detection limit of 20pg. We describe the spectrum of fatty-acyl chains in the cellular phosphoinositides. Consistent with previous work in other mammalian primary cells, the 38:4 fatty-acyl chains dominate in the phosphoinositides of the pineal gland and of superior cervical ganglia, and many additional fatty acid combinations are found at low abundance. However, Chinese hamster ovary cells and human embryonic kidney cells (tsA201) in culture have different fatty-acyl chain profiles that change with growth state. Their 38:4 lipids lose their dominance as cultures approach confluence. The method has good time resolution and follows well the depletion in <20s of both PIP2 and PIP that results from strong activation of Gq-coupled receptors. The receptor-activated phospholipase C exhibits no substrate selectivity among the various fatty-acyl chain combinations. •Refined quantitative mass spectrometry of cell phosphoinositides•Resolution of many fatty-acyl chain combinations in phosphoinositides•Dominance of 38:4 lipids in mammalian primary cells but not in cultured cells•Change of fatty-acyl profile with culture conditions•Fast kinetics (<10s) of phosphoinositide depletion after receptor activation
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	1388-1981 1879-2618
DOI:	10.1016/j.bbalip.2017.02.002