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Phosphoinositide Analysis by Liquid Chromatography–Mass Spectrometry

  • Trevor R. PettittEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 645)

Abstract

The phosphoinositides are a highly dynamic group of molecules implicated in many cellular control processes; however, the analysis of many of these structures has proven very difficult and time-consuming, with limited sensitivity and/or discrimination. Recent developments in LCMS now provide the prospect of routine structural and quantitative analysis of all the known phosphoinositides (and possibly some as yet unidentified structures) at high sensitivity in any biological sample. The procedures described here give very high extraction recovery from a variety of biological matrices and enable chromatographic resolution of most phosphoinositides as their native structures. When coupled with the accurate mass and fragmentation capabilities of an MS, full structural and isomeric identification can be achieved.

Key words

Lipids Phospholipids Phosphoinositides Phosphatidylinositol Lipidomics HPLC LCMS Mass spectrometry Normal-phase 

Lipid Abbreviations

PtdIns

phosphatidylinositol

PtdIns4P

phosphatidylinositol 4-phosphate

PtdIns3P

phosphatidylinositol 3-phosphate

PtdIns5P

phosphatidylinositol 5-phosphate

PtdIns(4,5)P2

phosphatidylinositol 4,5-bisphosphate

PtdIns(3,4)P2

phosphatidylinositol 3,4-bisphosphate

PtdIns(3,5)P2

phosphatidylinositol 3,5-bisphosphate

PtdIns(3,4,5)P3

phosphatidylinositol 3,4,5-trisphosphate

Notes

Acknowledgments

The author would like to thank Professor Michael Wakelam in whose laboratory much of this methodology was developed. This work was supported by grants from the Wellcome Trust.

References

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.CRUK Institute for Cancer StudiesUniversity of BirminghamBirminghamUK

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