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

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Kai Schuhmann - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Kristina Srzentić - , École Polytechnique Fédérale de Lausanne (Autor:in)
  • Konstantin O. Nagornov - , École Polytechnique Fédérale de Lausanne (Autor:in)
  • Henrik Thomas - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)
  • Theresia Gutmann - , Deutsches Zentrum für Diabetesforschung (DZD) e.V., Paul Langerhans Institut Dresden (PLID) des Helmholtz Zentrum München (Autor:in)
  • Ünal Coskun - , Deutsches Zentrum für Diabetesforschung (DZD) e.V., Paul Langerhans Institut Dresden (PLID) des Helmholtz Zentrum München (Autor:in)
  • Yury O. Tsybin - , École Polytechnique Fédérale de Lausanne (Autor:in)
  • Andrej Shevchenko - , Max Planck Institute of Molecular Cell Biology and Genetics (Autor:in)

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.

Details

OriginalspracheEnglisch
Seiten (von - bis)12857-12865
Seitenumfang9
FachzeitschriftAnalytical chemistry
Jahrgang89
Ausgabenummer23
PublikationsstatusVeröffentlicht - 5 Dez. 2017
Peer-Review-StatusJa
Extern publiziertJa

Externe IDs

Scopus 85037542286
PubMed 29111682
ORCID /0000-0003-4375-3144/work/142255267

Schlagworte

ASJC Scopus Sachgebiete