High throughput discovery of functional protein modifications by Hotspot Thermal Profiling.

Nature Methods
Jun X HuangRaymond E Moellering

Abstract

Mass spectrometry enables global analysis of posttranslationally modified proteoforms from biological samples, yet we still lack methods to systematically predict, or even prioritize, which modification sites may perturb protein function. Here we describe a proteomic method, Hotspot Thermal Profiling, to detect the effects of site-specific protein phosphorylation on the thermal stability of thousands of native proteins in live cells. This massively parallel biophysical assay unveiled shifts in overall protein stability in response to site-specific phosphorylation sites, as well as trends related to protein function and structure. This method can detect intrinsic changes to protein structure as well as extrinsic changes to protein-protein and protein-metabolite interactions resulting from phosphorylation. Finally, we show that functional 'hotspot' protein modification sites can be discovered and prioritized for study in a high-throughput and unbiased fashion. This approach is applicable to diverse organisms, cell types and posttranslational modifications.

References

Jun 11, 1999·Genes & Development·A C GingrasN Sonenberg
Dec 31, 2005·Nucleic Acids Research·Tzong-Yi LeeTzu-Hao Wang
Jun 6, 2006·Proteins·Chin-Sheng YuJenn-Kang Hwang
Dec 7, 2007·The Journal of Cell Biology·Jonathan D HumphriesChristoph Ballestrem
Mar 10, 2009·Current Opinion in Cell Biology·Tony Hunter
Aug 25, 2010·The Journal of Cell Biology·Tobias C Walther, Matthias Mann
Dec 25, 2010·Cell·Edward L HuttlinSteven P Gygi
Jan 11, 2011·Neurochemistry International·Ludovic MartinFaraj Terro
May 19, 2012·Nature Chemical Biology·Christina M AgapakisPamela A Silver
Jul 21, 2012·Nature Protocols·Morten KällbergJinbo Xu
Feb 28, 2013·Nature Methods·Lloyd M SmithUNKNOWN Consortium for Top Down Proteomics
Nov 5, 2013·Molecular & Cellular Proteomics : MCP·Jesper V Olsen, Matthias Mann
Oct 4, 2014·Science·Mikhail M SavitskiGerard Drewes
Jul 15, 2015·Proceedings of the National Academy of Sciences of the United States of America·Naotaka SekiyamaMélissa Léger-Abraham
Aug 19, 2015·Nature Biotechnology·Sean J HumphreyMatthias Mann
Sep 22, 2015·Nature Methods·Kilian V M HuberGiulio Superti-Furga
Feb 20, 2016·Molecular Systems Biology·Daria GawronPetra Van Damme
Apr 7, 2016·Annual Review of Analytical Chemistry·Forest M White, Alejandro Wolf-Yadlin
Jun 18, 2016·Analytical Chemistry·Jae Won ChangRaymond E Moellering
Sep 16, 2016·Nature·Ruedi Aebersold, Matthias Mann
May 13, 2017·Biophysical Journal·Kiavash GarakaniMohammad R K Mofrad
Mar 7, 2018·Molecular Systems Biology·Alireza AzimiGianluca Maddalo

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Citations

Mar 7, 2020·Molecular Systems Biology·André MateusMikhail M Savitski
Jun 4, 2020·Mass Spectrometry Reviews·Teck Yew LowMingliang Ye
Oct 22, 2020·Chemical Communications : Chem Comm·Fan Yang, Chu Wang
Nov 26, 2020·Developmental Cell·Chongde WuYansheng Liu
Jan 22, 2021·Journal of Proteome Research·Brendan M FloydEdward M Marcotte
Dec 11, 2020·Nature·André MateusMikhail M Savitski
Feb 13, 2021·Cell Chemical Biology·Louis P ConwayChristopher G Parker
Feb 17, 2021·Molecular Systems Biology·Joel SelkrigMikhail M Savitski
Feb 28, 2021·Nature Communications·Amir Ata SaeiRoman A Zubarev
Mar 9, 2020·Current Opinion in Chemical Biology·Jun X HuangRaymond E Moellering
Mar 4, 2021·Angewandte Chemie·Wang WanYu Liu
May 29, 2021·Cellular and Molecular Life Sciences : CMLS·Teck Yew LowChris Soon Heng Tan
Jun 9, 2021·Biochemical Society Transactions·Bernhard Hoermann, Maja Köhn
Jun 1, 2021·Frontiers in Molecular Biosciences·Carlos H Vieira-Vieira, Matthias Selbach
Jun 19, 2021·Nature Methods·Ian R SmithJudit Villén
Jun 19, 2021·Nature Methods·Clément M PotelMikhail M Savitski
Jun 19, 2021·Nature Methods·Jun X HuangRaymond E Moellering
Jul 1, 2021·Communications Biology·Siqi FangOliver M Crook
Jul 23, 2021·Molecular Systems Biology·Andre MateusIlaria Piazza
Oct 16, 2021·Expert Review of Proteomics·Xinyue LiuJoao A Paulo

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Methods Mentioned

BETA
X-ray
thermal shift
electrophoresis
confocal
PCR
transfection

Software Mentioned

R
RaptorX
PhosphoSite
modstat
trypstat
ImageJ
HTP
Integrated Proteomics Pipeline ( IP2 )
Prism5
Metamorph

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