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

Abstract:

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

Authors: K. Schuhmann, K. Srzentic, K. O. Nagornov, H. Thomas, T. Gutmann, U. Coskun, Y. O. Tsybin, A. Shevchenko

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Views: 4155

Created: 8th Jan 2018 at 10:39

Last updated: 8th Mar 2024 at 07:44

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