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Quantitative Analysis of Polyphosphoinositide, Bis(monoacylglycero)phosphate, and Phosphatidylglycerol Species by Shotgun Lipidomics After Methylation

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Mass Spectrometry-Based Lipidomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2306))

Abstract

Phospholipids play important roles in biological process even at a very low level. For example, bis(monoacylglycerol)phosphate (BMP) is involved in the pathogenesis of lysosomal storage diseases, and polyphosphoinositides (PPI) play critical roles in cellular signaling and functions. Phosphatidylglycerol (PG), a structural isomer of BMP, mediates lipid–protein and lipid–lipid interactions, and inhibits platelet activating factor and phosphatidylcholine transferring. However, due to their low abundance, the analysis of these phospholipids from biological samples is technically challenging. Therefore, the cellular function and metabolism of these phospholipids are still elusive. This chapter overviews a novel method of shotgun lipidomics after methylation with trimethylsilyl-diazomethane (TMS-D) for accurate and comprehensive analysis of these phospholipid species in biological samples. Firstly, a modified Bligh and Dyer procedure is performed to extract tissue lipids for PPI analysis, whereas modified methyl-tert-butylether (MTBE) extraction and modified Folch extraction methods are described to extract tissue lipids for PPI analysis. Secondly, TMS-D methylation is performed to derivatize PG/BMP and PPI, respectively. Then, we described the shotgun lipidomics strategies that can be used as cost-effective and relatively high-throughput methods to determine BMP, PG, and PPI species and isomers with different phosphate position(s) and fatty acyl chains. The described method of shotgun lipidomics after methylation achieves feasible and reliable quantitative analysis of low-abundance lipid classes. The application of this novel method should enable us to reveal the metabolism and functions of these phospholipids in healthy and disease states.

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Acknowledgments

This work was partially supported by National Institute of Aging Grant RF1 AG061872, National Institute of Neurological Disorders and Stroke Grant U54 NS110435, the institutional research funds from the University of Texas Health Science Center at San Antonio (UT Health SA), the Mass Spectrometry Core Facility at UT Health SA, and the Methodist Hospital Foundation.

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Authors and Affiliations

  1. Barshop Institute for Longevity and Aging Studies, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    Meixia Pan, Chao Qin & Xianlin Han

  2. Department of Medicine—Diabetes, University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    Xianlin Han

Authors
  1. Meixia Pan
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  2. Chao Qin
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  3. Xianlin Han
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Editors and Affiliations

  1. Mass Spectrometry Resource Division of Endocrinology Metabolism, and Lipid Research Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA

    Fong-Fu Hsu

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Pan, M., Qin, C., Han, X. (2021). Quantitative Analysis of Polyphosphoinositide, Bis(monoacylglycero)phosphate, and Phosphatidylglycerol Species by Shotgun Lipidomics After Methylation. In: Hsu, FF. (eds) Mass Spectrometry-Based Lipidomics. Methods in Molecular Biology, vol 2306. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1410-5_6

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  • DOI: https://doi.org/10.1007/978-1-0716-1410-5_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1409-9

  • Online ISBN: 978-1-0716-1410-5

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