title: Proteome scale turnover analysis in live animals using stable isotope metabolic labeling.
abstract: At present most quantitative proteomics investigations are focused on the analysis of protein expression differences between two or more sample specimens. With each analysis a static snapshot of a cellular state is captured with regard to protein expression. However, any information on protein turnover cannot be obtained using classic methodologies. Protein turnover, the result of protein synthesis and degradation, represents a dynamic process, which is of equal importance to understanding physiological processes. Methods employing isotopic tracers have been developed to measure protein turnover. However, applying these methods to live animals is often complicated by the fact that an assessment of precursor pool relative isotope abundance is required. Also, data analysis becomes difficult in case of low label incorporation, which results in a complex convolution of labeled and unlabeled peptide mass spectrometry signals. Here we present a protein turnover analysis method that circumvents this problem using a (15)N-labeled diet as an isotopic tracer. Mice were fed with the labeled diet for limited time periods and the resulting partially labeled proteins digested and subjected to tandem mass spectrometry. For the interpretation of the mass spectrometry data, we have developed the ProTurnyzer software that allows the determination of protein fractional synthesis rates without the need of precursor relative isotope abundance information. We present results validating ProTurnyzer with Escherichia coli protein data and apply the method to mouse brain and plasma proteomes for automated turnover studies.
authors.affiliation: Max Planck Institute of Psychiatry, Proteomics and Biomarkers, Munich, Germany.
authors.forename: Yaoyang
authors.initials: Y
authors.lastname: Zhang
authors.forename: Stefan
authors.initials: S
authors.lastname: Reckow
authors.forename: Christian
authors.initials: C
authors.lastname: Webhofer
authors.forename: Michael
authors.initials: M
authors.lastname: Boehme
authors.forename: Philipp
authors.initials: P
authors.lastname: Gormanns
authors.forename: Wolfgang M
authors.initials: WM
authors.lastname: Egge-Jacobsen
authors.forename: Christoph W
authors.initials: CW
authors.lastname: Turck
mesh_terms: D000465:Algorithms; D000818:Animals; D007554:Isotopes; D008297:Male; D051379:Mice; D008811:Mice, Inbred DBA; D011506:Proteins / Q000378:metabolism; D020543:Proteome
chemical_list: D007554:Isotopes; D011506:Proteins; D020543:Proteome