title: Monitoring PPARG-Induced Changes in Glycolysis by Selected Reaction Monitoring Mass Spectrometry.
abstract: As cells develop and differentiate, they change in function and morphology, which often precede earlier changes in signaling and metabolic control. Here we present a selected reaction monitoring (SRM) approach which allows for the parallel quantification of metabolic regulators and their downstream targets.In particular we explain and describe how to monitor abundance changes of glycolytic enzymes upon PPARγ activation by using a label-free or a stable isotope-labeled standard peptide (SIS peptides) approach applying triple-quadrupole mass spectrometry. We further outline how to fractionate the cell lysate into cytosolic and nuclear fractions to enhance the sensitivity of the measurements and to investigate the dynamic concentration changes in those compartments.
authors.affiliation: Leibniz-Institut für Analytische Wissenschaften-ISAS-e.V., 44227, Dortmund, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hentschel
authors.affiliation: Leibniz-Institut für Analytische Wissenschaften-ISAS-e.V., 44227, Dortmund, Germany. robert.ahrends@isas.de.
authors.forename: Robert
authors.initials: R
authors.lastname: Ahrends
mesh_terms: D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D006019:Glycolysis; D006801:Humans; D013058:Mass Spectrometry; D047495:PPAR gamma / Q000378:metabolism*; D010455:Peptides; D020543:Proteome; D040901:Proteomics / Q000379:methods*
chemical_list: D047495:PPAR gamma; D010455:Peptides; D020543:Proteome
keywords: Adipocytes; Mass spectrometry; Metabolic control; Proteomics; Quantification; SIS peptides; Selected reaction monitoring