A quantitative mass spectrometry-based approach to monitor the dynamics of endogenous chromatin-associated protein complexes

被引:0
作者
Evangelia K. Papachristou
Kamal Kishore
Andrew N. Holding
Kate Harvey
Theodoros I. Roumeliotis
Chandra Sekhar Reddy Chilamakuri
Soleilmane Omarjee
Kee Ming Chia
Alex Swarbrick
Elgene Lim
Florian Markowetz
Matthew Eldridge
Rasmus Siersbaek
Clive S. D’Santos
Jason S. Carroll
机构
[1] Cancer Research UK Cambridge Institute,
[2] University of Cambridge,undefined
[3] Garvan Institute of Medical Research,undefined
[4] Darlinghurst,undefined
[5] Wellcome Trust Sanger Institute,undefined
[6] St Vincent’s Clinical School,undefined
[7] UNSW,undefined
来源
Nature Communications | / 9卷
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摘要
Understanding the dynamics of endogenous protein–protein interactions in complex networks is pivotal in deciphering disease mechanisms. To enable the in-depth analysis of protein interactions in chromatin-associated protein complexes, we have previously developed a method termed RIME (Rapid Immunoprecipitation Mass spectrometry of Endogenous proteins). Here, we present a quantitative multiplexed method (qPLEX-RIME), which integrates RIME with isobaric labelling and tribrid mass spectrometry for the study of protein interactome dynamics in a quantitative fashion with increased sensitivity. Using the qPLEX-RIME method, we delineate the temporal changes of the Estrogen Receptor alpha (ERα) interactome in breast cancer cells treated with 4-hydroxytamoxifen. Furthermore, we identify endogenous ERα-associated proteins in human Patient-Derived Xenograft tumours and in primary human breast cancer clinical tissue. Our results demonstrate that the combination of RIME with isobaric labelling offers a powerful tool for the in-depth and quantitative characterisation of protein interactome dynamics, which is applicable to clinical samples.
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