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Anal Chem. 2017 Dec 5;89(23):12857-12865. doi: 10.1021/acs.analchem.7b03437. Epub 2017 Nov 22.

Monitoring Membrane Lipidome Turnover by Metabolic 15N Labeling and Shotgun Ultra-High-Resolution Orbitrap Fourier Transform Mass Spectrometry.

Author information

1
Max Planck Institute of Molecular Cell Biology and Genetics , Pfotenhauerstrasse 108, 01307 Dresden, Germany.
2
Ecole Polytechnique Fédérale de Lausanne , 1015 Lausanne, Switzerland.
3
Spectroswiss , EPFL Innovation Park, Building I, 1015 Lausanne, Switzerland.
4
Paul Langerhans Institute Dresden of the Helmholtz Zentrum München at the University Hospital and Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden , Fetscher Strasse 74, 01307 Dresden, Germany.
5
German Center for Diabetes Research , Ingolstädter Landstrasse 1, 85764 Neuherberg, Germany.

Abstract

Lipidomes undergo permanent extensive remodeling, but how the turnover rate differs between lipid classes and molecular species is poorly understood. We employed metabolic 15N 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 × 106 (m/z 200) baseline resolved peaks of 13C isotopes of unlabeled and monoisotopic peaks of 15N labeled lipids (Δm = 0.0063 Da). Therefore, the rate of metabolic 15N 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.

PMID:
29111682
DOI:
10.1021/acs.analchem.7b03437
[Indexed for MEDLINE]

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