Monitoring Membrane Lipidome Turnover by Metabolic (15)N Labeling and Shotgun Ultra-High-Resolution Orbitrap Fourier Transform Mass Spectrometry.
Lipidomes undergo permanent extensive remodeling, but how the turnover rate differs between lipid classes and molecular species is poorly understood. We employed metabolic (15)N labeling and shotgun ultra-high-resolution mass spectrometry (sUHR) to quantify the absolute (molar) abundance and determine the turnover rate of glycerophospholipids and sphingolipids by direct analysis of total lipid extracts. sUHR performed on a commercial Orbitrap Elite instrument at the mass resolution of 1.35 x 10(6) (m/z 200) baseline resolved peaks of (13)C isotopes of unlabeled and monoisotopic peaks of (15)N labeled lipids (Deltam = 0.0063 Da). Therefore, the rate of metabolic (15)N labeling of individual lipid species could be determined without compromising the scope, accuracy, and dynamic range of full-lipidome quantitative shotgun profiling. As a proof of concept, we employed sUHR to determine the lipidome composition and fluxes of 62 nitrogen-containing membrane lipids in human hepatoma HepG2 cells.
SEEK ID: https://seek.lisym.org/publications/49
PubMed ID: 29111682
Projects: LiSyM Pillar I: Early Metabolic Injury (LiSyM-EMI)
Publication type: Not specified
Journal: Anal Chem
Citation: Anal Chem. 2017 Dec 5;89(23):12857-12865. doi: 10.1021/acs.analchem.7b03437. Epub 2017 Nov 22.
Date Published: 5th Dec 2017
Registered Mode: Not specified
Views: 4155
Created: 8th Jan 2018 at 10:39
Last updated: 8th Mar 2024 at 07:44
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