Affinity proteomics to study endogenous protein complexes: pointers, pitfalls, preferences and perspectives

BioTechniques
John LaCavaMichael P Rout

Abstract

Dissecting and studying cellular systems requires the ability to specifically isolate distinct proteins along with the co-assembled constituents of their associated complexes. Affinity capture techniques leverage high affinity, high specificity reagents to target and capture proteins of interest along with specifically associated proteins from cell extracts. Affinity capture coupled to mass spectrometry (MS)-based proteomic analyses has enabled the isolation and characterization of a wide range of endogenous protein complexes. Here, we outline effective procedures for the affinity capture of protein complexes, highlighting best practices and common pitfalls.

References

Nov 1, 1977·Nucleic Acids Research·H J LiC Chang
May 2, 1986·European Journal of Biochemistry·T MoksM Uhlén
Feb 1, 1987·Protein Engineering·B NilssonM Uhlén
Aug 12, 1983·Journal of Immunological Methods·R LindmarkJ Sjöquist
Dec 24, 1996·Proceedings of the National Academy of Sciences of the United States of America·D FirsovB C Rossier
Aug 19, 1997·Proceedings of the National Academy of Sciences of the United States of America·M C SchultzB A Altheim
Oct 31, 1997·Journal of Molecular Biology·M TashiroG T Montelione
Mar 26, 1998·FEMS Immunology and Medical Microbiology·B JanssonP A Nygren
Feb 3, 1999·Annual Review of Genetics·J W Jarvik, C A Telmer
Mar 17, 1999·Methods : a Companion to Methods in Enzymology·M C Schultz
Jun 9, 1999·Proceedings of the National Academy of Sciences of the United States of America·Y OdaB T Chait
Sep 24, 1999·Protein Science : a Publication of the Protein Society·C Q Pan, R A Lazarus
Feb 26, 2000·Science·W L DeLanoJ A Wells
Apr 19, 2000·BioTechniques·R HernanB Brizzard
Apr 29, 2000·Electrophoresis·K Gevaert, J Vandekerckhove
Nov 15, 2000·Genome Research·J L HartleyM A Brasch
Oct 9, 2001·Trends in Biochemical Sciences·R J Ellis
Jun 21, 2002·Methods in Enzymology·Marie E Petracek, Mark S Longtine
Sep 21, 2002·Molecular and Cellular Biology·Nevan J KroganJack F Greenblatt
Mar 14, 2003·Nature·Ruedi Aebersold, Matthias Mann
Jul 31, 2003·Biotechnology and Bioengineering·Beat P KramerMartin Fussenegger
Oct 17, 2003·Nature·Won-Ki HuhErin K O'Shea
Oct 17, 2003·Nature·Sina GhaemmaghamiJonathan S Weissman
Jun 3, 2004·Electrophoresis·Giovanni CandianoPier Giorgio Righetti
Sep 25, 2004·Genetic Engineering·Christian Berens, Wolfgang Hillen
Oct 19, 2004·Genome Research·Daniela S GerhardUNKNOWN MGC Project Team
Nov 3, 2004·Biochimica Et Biophysica Acta·Annela M SeddonPaula J Booth
Nov 17, 2004·Langmuir : the ACS Journal of Surfaces and Colloids·Martin LundqvistBengt-Harald Jonsson
Jan 13, 2005·Science's STKE : Signal Transduction Knowledge Environment·Iain M Cheeseman, Arshad Desai

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Citations

Jan 27, 2017·Molecular BioSystems·Spencer HavisSteven J Bark
Sep 24, 2015·PloS One·Md Babul HossainToshinobu Tokumoto
Jan 24, 2019·Cellular and Molecular Life Sciences : CMLS·Yinxing MaSvetlana Dokudovskaya
May 6, 2015·Nature Methods·Zhanna HakhverdyanJohn LaCava
Mar 17, 2019·The Journal of Biological Chemistry·Rajaiah PerguSivaram V S Mylavarapu
Nov 12, 2017·Scientific Reports·Yinxing MaSvetlana Dokudovskaya
May 20, 2020·Memórias do Instituto Oswaldo Cruz·Yirys Díaz-OlmosFabricio K Marchini
Sep 23, 2020·Nucleic Acids Research·Yuhui DouJohn LaCava
Nov 30, 2019·Journal of Cell Science·Jian XiongGuangwei Du
Apr 16, 2016·Journal of the American Society for Mass Spectrometry·Brian T ChaitYi Shi
Oct 21, 2016·Journal of Proteome Research·Rodolfo CiuffaRuedi Aebersold
Jul 13, 2016·RNA·Michal DomanskiJohn LaCava

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Methods Mentioned

BETA
affinity capture
affinity
immunoprecipitation
tandem affinity purification
transfections
transfection
FACS
light microscopy
DNA shearing
to affinity

Software Mentioned

Retsch Cryomill
Gateway

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