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

作者：Papachristou Evangelia K; Kishore Kamal; Holding Andrew N; Harvey Kate; Roumeliotis Theodoros I; Chilamakuri Chandra Sekhar Reddy; Omarjee Soleilmane; Chia Kee Ming; Swarbrick Alex; Lim Elgene; Markowetz Florian; Eldridge Matthew; Sier**aek Rasmus^*; D'Santos Clive S^*; Carroll Jason S^*

来源：Nature Communications, 2018, 9(1): 2311.

DOI：10.1038/s41467-018-04619-5

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 (ER alpha) interactome in breast cancer cells treated with 4-hydroxytamoxifen. Furthermore, we identify endogenous ER alpha-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.

• 出版日期2018-6-13
• 单位河北医科大学