Chapter 24
Targeted Lipidomics of Mycobacterial Lipids and Glycolipids
Emilie Layre
Abstract
Decades of study have highlighted the richness and uniqueness of the repertoire of lipid and glycolipid
families produced by mycobacteria. Many of these families potently regulate host immune responses, in
stimulatory or suppressive ways. Thus, the global study of this repertoire in different genetic backgrounds
or under model conditions of infection is gaining interest. Despite the difficulties associated with the
specificities of this repertoire, the field of mass spectrometry-based lipidomics of mycobacteria has recently
made considerable progress, particularly at the analytical level. There is still considerable scope for further
progress, especially with regard to the development of an efficient bioinfomatics pipeline for the analysis of
the large datasets generated. This chapter describes an HPLC-MS methodology allowing the simultaneous
screening of more than 20 of the lipid families produced by mycobacteria and provides recommendations to
analyze the generated data given the state-of-the-art.
Key words Mycobacterium, Lipidomics, Mass spectrometry, HPLC-MS
1 Introduction
Different analytical techniques are used to analyze lipids and glycolipids including thin layer chromatography (TLC), nuclear magnetic resonance (NMR), and mass spectrometry (MS), which have
their strengths and weaknesses, as previously reviewed [1]. Nevertheless, if by lipidomic we mean the rapid and “global” analysis of
the lipids and glycolipids that characterize a given system, mass
spectrometry appears to be the method of choice because it provides sensitivity, resolution, and high throughput and allows assessing the acyl-form diversity of the lipid families.
Lipidomics approaches, which have benefited from the enormous progress of mass spectrometry, are today diverse and highly
developed, in particular, for the analysis of eukaryotic lipids
[2]. These approaches share main steps: (1) sample preparation
(lipid extraction), (2) analytical measurement (MS-based detection), (3) data processing (lipid identification and quantification),
and (4) data interpretation, which depend on biological questions
and rely on more or less complex bioinformatics pipelines. The type
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_24, © Springer Science+Business Media, LLC, part of Springer Nature 2021
549
Targeted Lipidomics of Mycobacterial Lipids and Glycolipids
Emilie Layre
Abstract
Decades of study have highlighted the richness and uniqueness of the repertoire of lipid and glycolipid
families produced by mycobacteria. Many of these families potently regulate host immune responses, in
stimulatory or suppressive ways. Thus, the global study of this repertoire in different genetic backgrounds
or under model conditions of infection is gaining interest. Despite the difficulties associated with the
specificities of this repertoire, the field of mass spectrometry-based lipidomics of mycobacteria has recently
made considerable progress, particularly at the analytical level. There is still considerable scope for further
progress, especially with regard to the development of an efficient bioinfomatics pipeline for the analysis of
the large datasets generated. This chapter describes an HPLC-MS methodology allowing the simultaneous
screening of more than 20 of the lipid families produced by mycobacteria and provides recommendations to
analyze the generated data given the state-of-the-art.
Key words Mycobacterium, Lipidomics, Mass spectrometry, HPLC-MS
1 Introduction
Different analytical techniques are used to analyze lipids and glycolipids including thin layer chromatography (TLC), nuclear magnetic resonance (NMR), and mass spectrometry (MS), which have
their strengths and weaknesses, as previously reviewed [1]. Nevertheless, if by lipidomic we mean the rapid and “global” analysis of
the lipids and glycolipids that characterize a given system, mass
spectrometry appears to be the method of choice because it provides sensitivity, resolution, and high throughput and allows assessing the acyl-form diversity of the lipid families.
Lipidomics approaches, which have benefited from the enormous progress of mass spectrometry, are today diverse and highly
developed, in particular, for the analysis of eukaryotic lipids
[2]. These approaches share main steps: (1) sample preparation
(lipid extraction), (2) analytical measurement (MS-based detection), (3) data processing (lipid identification and quantification),
and (4) data interpretation, which depend on biological questions
and rely on more or less complex bioinformatics pipelines. The type
Tanya Parish and Anuradha Kumar (eds.), Mycobacteria Protocols, Methods in Molecular Biology, vol. 2314,
https://doi.org/10.1007/978-1-0716-1460-0_24, © Springer Science+Business Media, LLC, part of Springer Nature 2021
549