M-Track: detecting short-lived protein-protein interactions in vivo.

Nature Methods
Aurora ZuzuarreguiEgon Ogris

Abstract

We developed a protein-proximity assay in yeast based on fusing a histone lysine methyltransferase onto a bait and its substrate onto a prey. Upon binding, the prey is stably methylated and detected by methylation-specific antibodies. We applied this approach to detect varying interaction affinities among proteins in a mitogen-activated protein kinase pathway and to detect short-lived interactions between protein phosphatase 2A and its substrates that have so far escaped direct detection.

References

Jun 22, 2000·Yeast·V V Kushnirov
Feb 21, 2004·Methods in Enzymology·Laura Perez-BurgosThomas Jenuwein
Feb 19, 2005·Current Opinion in Structural Biology·Jacob Piehler
Jun 28, 2008·Journal of the American Chemical Society·Marta Fernández-SuárezAlice Y Ting
Jan 27, 2010·Nature Methods·Mark F CiaccioRichard B Jones
Nov 13, 2010·Nucleic Acids Research·Chris StarkMike Tyers

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Citations

Aug 14, 2012·The EMBO Journal·Claudine KraftMatthias Peter
Aug 15, 2013·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Anna Jean Wirth, Martin Gruebele
Aug 11, 2015·ELife·Xiaoji SunAndreas Hochwagen
May 17, 2015·Nature Chemical Biology·Zhong YaoIgor Stagljar
Feb 18, 2015·Biochimica Et Biophysica Acta·Aurora ZuzuarreguiPaula Alepuz
Jan 21, 2014·Molecular Cell·Daniel PapinskiClaudine Kraft
Oct 5, 2016·ACS Chemical Biology·Sara FahsMaja Köhn
Nov 24, 2016·ELife·Dorotea FracchiollaSascha Martens
Mar 9, 2017·Science Signaling·Natalie RomanovWolfgang Reiter
Jul 14, 2017·Cells·Raffaela TorgglerClaudine Kraft
Jun 30, 2018·Mass Spectrometry Reviews·Kevin TitecaSven Eyckerman
Jul 19, 2019·The Journal of Cell Biology·Frank van DrogenMatthias Peter
Oct 28, 2019·Journal of Cell Science·David M HollensteinClaudine Kraft
Jun 19, 2019·Cell Communication and Signaling : CCS·Marion JanschitzWolfgang Reiter
Sep 25, 2021·EMBO Reports·David Maria HollensteinWolfgang Reiter

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Methods Mentioned

BETA
protein
phosphatase assay
immunoprecipitation
X-ray
dot blot
ELISA

Software Mentioned

Office PowerPoint
Track
Adobe Photoshop
ImageQuant
Adobe Acrobat

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