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

• DOI: 10.1038/s41467-018-04619-5
• Publication Date: 2018-06-07T09:50:58Z
• Authors: Evangelia K. Papa...; Kamal Kishore; Andrew N. Holding; Kate Harvey; Theodoros I. Roum...; Chandra Sekhar Re...; Soleilmane Omarjee; Kee Ming Chia; Alex Swarbrick; Elgene Lim; Florian Markowetz; Matthew Eldridge; Rasmus Siersbaek; Clive S. D’Santos; Jason S. Carroll
• Keywords: Interactome; Hard rime; Immunoprecipitation

Abstract Understanding the dynamics of endogenous protein–protein interactions in complex networks is pivotal deciphering disease mechanisms. To enable in-depth analysis protein chromatin-associated complexes, we have previously developed a method termed RIME (Rapid Immunoprecipitation Mass spectrometry Endogenous proteins). Here, present quantitative multiplexed (qPLEX-RIME), which integrates with isobaric labelling and tribrid mass for study interactome fashion increased sensitivity. Using qPLEX-RIME method, delineate temporal changes Estrogen Receptor alpha (ERα) breast cancer cells treated 4-hydroxytamoxifen. Furthermore, identify ERα-associated proteins human Patient-Derived Xenograft tumours primary clinical tissue. Our results demonstrate that combination offers powerful tool characterisation dynamics, applicable to samples.