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

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

*Kontaktforfatter

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

69 Downloads (Pure)

Abstract

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.

OriginalsprogEngelsk
Artikelnummer2311
TidsskriftNature Communications
Vol/bind9
Udgave nummer1
ISSN2041-1723
DOI
StatusUdgivet - 1. dec. 2018
Udgivet eksterntJa

Fingeraftryk

Dyk ned i forskningsemnerne om 'A quantitative mass spectrometry-based approach to monitor the dynamics of endogenous chromatin-associated protein complexes'. Sammen danner de et unikt fingeraftryk.

Citationsformater