A quantitative mass spectrometry-based approach to monitor the dynamics of endogenous chromatin-associated protein complexes.
View / Open Files
Authors
Harvey, Kate
Roumeliotis, Theodoros I
Chilamakuri, Chandra
Omarjee, Soleilmane
Chia, Kee Ming
Swarbrick, Alex
Lim, Elgene
Siersbaek, Rasmus
D'Santos, Clive S
Publication Date
2018-06-13Journal Title
Nature Communications
ISSN
2041-1723
Publisher
Springer Nature
Volume
9
Issue
1
Number
2311
Pages
2311-2311
Language
eng
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Papachristou, E., Kishore, K., Holding, A., Harvey, K., Roumeliotis, T. I., Chilamakuri, C., Omarjee, S., et al. (2018). A quantitative mass spectrometry-based approach to monitor the dynamics of endogenous chromatin-associated protein complexes.. Nature Communications, 9 (1. 2311), 2311-2311. https://doi.org/10.1038/s41467-018-04619-5
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.
Keywords
Breast cancer, Immunoprecipitation, Mass spectrometry, Protein–protein interaction networks
Sponsorship
Wellcome Trust (108467/Z/15/Z)
Cancer Research UK (C14303/A17197)
Breast Cancer Campaign (2012NovemberPR042)
European Research Council (646876)
Cancer Research UK (CRUK-A19274)
Identifiers
External DOI: https://doi.org/10.1038/s41467-018-04619-5
This record's URL: https://www.repository.cam.ac.uk/handle/1810/280762
Statistics
Total file downloads (since January 2020). For more information on metrics see the
IRUS guide.
Recommended or similar items
The following licence files are associated with this item: