Effective Method for Accurate and Sensitive Quantitation of Rapid Changes of Newly Synthesized Proteins.

Analytical Chemistry
Ming TongRonghu Wu

Abstract

Protein synthesis is quickly and tightly regulated in cells to adapt to the ever-changing extracellular and intracellular environment. Accurate quantitation of rapid protein synthesis changes can provide insights into protein functions and cellular activities, but it is very challenging to achieve because of the lack of effective analysis methods. Here, we developed an effective mass spectrometry-based method named quantitative O-propargyl-puromycin tagging (QOT) by integrating O-propargyl-puromycin (OPP) labeling, bioorthogonal chemistry, and multiplexed proteomics for global and quantitative analysis of rapid protein synthesis. The current method enables us to accurately quantitate rapid changes of newly synthesized proteins because, unlike amino acids and their analogs, OPP can be utilized by the ribosome immediately without being activated and conjugated to tRNA, and thus cell starvation or pretreatment is not required. This method was applied to quantitate rapid changes of protein synthesis in THP-1 macrophages treated with lipopolysaccharide (LPS). For 15-min labeling, >3000 proteins were quantitated, and the synthesis of 238 proteins was significantly altered, including transcription factors and cytokines. The results de...Continue Reading

References

Apr 2, 1996·Proceedings of the National Academy of Sciences of the United States of America·J HambletonA L DeFranco
Sep 7, 2000·The Journal of Immunology : Official Journal of the American Association of Immunologists·M G ScottR E Hancock
Oct 18, 2000·The Journal of Immunology : Official Journal of the American Association of Immunologists·E SchutyserJ Van Damme
Sep 18, 2002·The EMBO Journal·Aristides G EliopoulosPhilip N Tsichlis
Mar 13, 2003·Journal of the American Chemical Society·Qian WangM G Finn
Nov 12, 2005·Science·Rachael Z MurrayJennifer L Stow
Apr 6, 2006·Proceedings of the National Academy of Sciences of the United States of America·Joerg SchreiberRichard A Young
Jun 14, 2006·Proceedings of the National Academy of Sciences of the United States of America·Daniela C DieterichErin M Schuman
Sep 23, 2006·Physiology·Xuemin Wang, Christopher G Proud
Jan 16, 2007·BMC Immunology·Omar SharifNeil D Perkins
Dec 29, 2007·The Journal of Pathology·J R Bradley
Jun 5, 2008·The Journal of Immunology : Official Journal of the American Association of Immunologists·Yanyan WangBruce H Horwitz
Nov 19, 2008·Nature Reviews. Genetics·Zhong WangMichael Snyder
Mar 10, 2009·Immunobiology·Jennifer L StowDaniele Sangermani
Feb 27, 2010·Critical Reviews in Biochemistry and Molecular Biology·Monika Gooz
Jun 10, 2010·Molecular Systems Biology·Gabriele WeintzRoland Lang
Nov 26, 2010·The Journal of Biological Chemistry·Xuesong QianJianguo Liu
Dec 29, 2010·Cell Research·Michael J Morgan, Zheng-gang Liu
May 20, 2011·Nature·Björn SchwanhäusserMatthias Selbach
Jun 28, 2011·Immunopharmacology and Immunotoxicology·Haeyong LeeYoosik Yoon
Dec 14, 2011·Proceedings of the National Academy of Sciences of the United States of America·Jing LiuAdrian Salic
Apr 13, 2012·Journal of Leukocyte Biology·Gang PeiMaximiliano Gabriel Gutierrez
Jun 16, 2012·Cold Spring Harbor Perspectives in Biology·Adam M Benham
Jun 23, 2012·The Journal of Immunology : Official Journal of the American Association of Immunologists·Prachi P SinghJeffery S Schorey
Mar 12, 2013·Nature Methods·Andrew J M HowdenOreste Acuto
Apr 16, 2013·Biochimica Et Biophysica Acta·Markus Aebi
Jul 23, 2013·The Journal of Immunology : Official Journal of the American Association of Immunologists·Lydia HanssenUte Raffetseder
Nov 1, 1994·Journal of the American Society for Mass Spectrometry·J K EngJ R Yates
Jan 8, 2014·Molecular & Cellular Proteomics : MCP·Katrin Eichelbaum, Jeroen Krijgsveld
Jan 29, 2014·Nature Reviews. Genetics·Nicholas T Ingolia
Mar 29, 2014·Nature·Robert A J SignerSean J Morrison
Nov 12, 2014·Frontiers in Immunology·Rachael Zoe Murray, Jennifer Lea Stow
Mar 19, 2015·Annual Review of Biochemistry·Johnathan Labbadia, Richard I Morimoto

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Citations

Aug 9, 2021·Current Opinion in Chemical Biology·Wouter van BergenMarc P Baggelaar
Nov 23, 2021·Pharmacological Research : the Official Journal of the Italian Pharmacological Society·Anwen YinLingmei Qian

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