Quantitative proteomics using 16O/18O labeling and linear ion trap mass spectrometry

Proteomics
Daniel López-FerrerJesús Vázquez

Abstract

Quantitative proteomics using stable isotopic 16O/18O labeling has emerged as a very powerful tool, since it has a number of advantages over other methods, including the simplicity of chemistry, the constant mass tag at the C termini and its general applicability. However, due to the small mass difference between labeled and unlabeled peptide species, this approach has usually been restricted to high-resolution mass spectrometers. In this study we explored whether the high-resolution scanning mode, together with the extremely high scanning speed of the linear IT allows the 16O/18O-labeling method to be used for accurate, large-scale quantitative analysis of proteomes. A protocol, including digestion, desalting, labeling, MS and quantitative analysis was developed and tested using protein standards and whole proteome extracts. Using this method we were able to identify and quantify 140 proteins from only 10 mug of a proteome extract from mesenchymal stem cells. Relative expression changes larger than twofold can be identified with this method at the 95% confidence level. Our results demonstrate that accurate quantitative analysis using 16O/18O labeling can be performed in the practice using linear IT MS, without compromising lar...Continue Reading

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Citations

Apr 8, 2011·Journal of the American Society for Mass Spectrometry·Qi Zhu, Tomasz Burzykowski
Jan 21, 2011·Journal of Proteome Research·Alexey V NefedovRovshan G Sadygov
Aug 9, 2008·Journal of Proteome Research·Daniel López-FerrerRichard D Smith
Aug 2, 2006·Molecular BioSystems·Jeffrey C Smith, Daniel Figeys
Sep 3, 2010·Molecular & Cellular Proteomics : MCP·Elena Bonzon-KulichenkoJesús Vázquez
Jun 1, 2012·Molecular & Cellular Proteomics : MCP·Pablo Martínez-AcedoJesús Vázquez
Jan 20, 2009·Briefings in Functional Genomics & Proteomics·Xiaoying YeJosip Blonder
Feb 10, 2012·Cold Spring Harbor Perspectives in Biology·Dennis van HoofChristine Mummery
Oct 11, 2008·Analytical and Bioanalytical Chemistry·Elmar LangenfeldKatrin Marcus
Jun 14, 2013·Critical Reviews in Clinical Laboratory Sciences·Daniela CretuVinod Chandran
Aug 11, 2015·Proteomics. Clinical Applications·Frances ZenónHoracio Serrano
Feb 6, 2007·Journal of Chromatography. a·Benito CañasJose Manuel Gallardo
Apr 8, 2016·Cell Reports·Adela GuarásJose A Enríquez
Sep 17, 2011·Journal of Proteomics·Elena Bonzon-KulichenkoJesús Vázquez
Jun 19, 2013·Journal of Proteomics·Marta MendesCelso Cunha
Jan 18, 2014·Journal of Molecular and Cellular Cardiology·Marisol Ruiz-MeanaDavid Garcia-Dorado
Dec 19, 2014·Cell Death & Disease·C Fernandez-SanzD Garcia-Dorado
Feb 12, 2014·Journal of Proteome Research·Pedro NavarroJesús Vázquez
Feb 12, 2013·Progress in Retinal and Eye Research·Gülgün Tezel
Jan 29, 2013·Biomedit︠s︡inskai︠a︡ khimii︠a︡·N E Moskaleva, V G Zgoda
Dec 2, 2020·Foods·Iciar MartinezMónica Carrera
Feb 27, 2007·Molecular & Cellular Proteomics : MCP·Antonio Ramos-FernándezJesús Vázquez

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