Lipidomics in Cell Biology

Jiménez‐Rojo, Noemi; Vacca, Fabrizio; Riezman, Howard

doi:10.1002/9783527836512.ch26

“…A DIA acquisition, designed in the Thermo Xcalibur method editor, was used to collect MS 1 , MS 2 and MS 3 (CID/OzID) scans over a 35 min duration with the MS 3 sn- isomer data for PC and PE species taking ∼20 min (the only data used here for sn -isomer characterisation). MS 1 spectra were collected for ca .…”

Section: Methods Cell Culturementioning

confidence: 99%

“…A DIA acquisition, designed in the Thermo Xcalibur method editor, was used to collect MS 1 , MS 2 and MS 3 (CID/OzID) scans over a 35 min duration with the MS 3 sn- isomer data for PC and PE species taking ∼20 min (the only data used here for sn -isomer characterisation). MS 1 spectra were collected for ca . 8 minutes with an m/z range of 350-1600 at a target mass resolution of 500,000 at m/z 200 (FWHM) and an automatic gain control (AGC) at 50% (manufacturer units) and a 50 ms maximum injection time.…”

Section: Methods Cell Culturementioning

confidence: 99%

“…Glycerophospholipids (GPL) are one of the most ubiquitous lipid classes in biology and the most abundant in vertebrata. GPLs play many important functions within cellular membranes, including regulating membrane mechanics, signalling and protein function 1-3 , whilst changes in lipid metabolism and the lipidome are a hallmark of many diseases, including cancers 4, 5 . The biological function and metabolic origin of lipids depends on their precise chemical structure, including headgroup (defining the GPL class), the length and degree of unsaturation of each acyl chain, and the positions of each chain on the glycerol backbone (defined as the stereospecific numbering [ sn ]-position) 6, 7 .…”

Section: Mainmentioning

confidence: 99%

See 1 more Smart Citation

A High-Throughput Data-Independent Acquisition Workflow for Deep Characterisation of thesn-Isomer Lipidome

Michael,

Young,

Balez

et al. 2023

Preprint

0

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We report a workflow based on ozone-induced dissociation for untargeted characterization of hundreds ofsn-resolved glycerophospholipid isomers from biological extracts in under 20 minutes, coupled with an automated data analysis pipeline. It provides an order of magnitude increase in the number ofsn-isomer pairs identified compared to previous reports, reveals thatsn-isomer populations are tightly regulated and significantly different between cell lines, and enables identification of rare lipids containing ultra-long chain monounsaturated acyl chains.

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Deep Characterisation of the sn‐Isomer Lipidome Using High‐Throughput Data‐Independent Acquisition and Ozone‐Induced Dissociation**

Michael,

Young,

Balez

et al. 2024

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In recent years there has been a significant interest in the development of innovative lipidomics techniques capable of resolving lipid isomers. To date, methods applied to resolving sn‐isomers have resolved only a limited number of species. We report a workflow based on ozone‐induced dissociation for untargeted characterisation of hundreds of sn‐resolved glycerophospholipid isomers from biological extracts in under 20 minutes, coupled with an automated data analysis pipeline. It provides an order of magnitude increase over the number of sn‐isomer pairs identified compared to previous reports and reveals that sn‐isomer populations are tightly regulated and significantly different between cell lines. The sensitivity of this method and potential for de novo molecular discovery is further demonstrated by the identification of unexpected lipids containing ultra‐long chain monounsaturated acyl chains at the sn‐1 position.

show abstract

Deep Characterisation of the sn‐Isomer Lipidome Using High‐Throughput Data‐Independent Acquisition and Ozone‐Induced Dissociation**

Michael,

Young,

Balez

et al. 2024

Angew Chem Int Ed

3

0

View full text Add to dashboard Cite

In recent years there has been a significant interest in the development of innovative lipidomics techniques capable of resolving lipid isomers. To date, methods applied to resolving sn‐isomers have resolved only a limited number of species. We report a workflow based on ozone‐induced dissociation for untargeted characterisation of hundreds of sn‐resolved glycerophospholipid isomers from biological extracts in under 20 minutes, coupled with an automated data analysis pipeline. It provides an order of magnitude increase over the number of sn‐isomer pairs identified compared to previous reports and reveals that sn‐isomer populations are tightly regulated and significantly different between cell lines. The sensitivity of this method and potential for de novo molecular discovery is further demonstrated by the identification of unexpected lipids containing ultra‐long chain monounsaturated acyl chains at the sn‐1 position.

show abstract

Lipidomics in Cell Biology

Cited by 3 publications

References 97 publications

A High-Throughput Data-Independent Acquisition Workflow for Deep Characterisation of thesn-Isomer Lipidome

A High-Throughput Data-Independent Acquisition Workflow for Deep Characterisation of thesn-Isomer Lipidome

Deep Characterisation of the sn‐Isomer Lipidome Using High‐Throughput Data‐Independent Acquisition and Ozone‐Induced Dissociation**

Deep Characterisation of the sn‐Isomer Lipidome Using High‐Throughput Data‐Independent Acquisition and Ozone‐Induced Dissociation**

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