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Tvardovskiy, A. ; Nguyen, N.* ; Bartke, T.

Identifying specific protein interactors of nucleosomes carrying methylated histones using quantitative mass spectrometry.

In: Histone Methyltransferases. Berlin [u.a.]: Springer, 2022. 327-403 (Methods Mol. Biol. ; 2529)
DOI PMC
Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
Chemical modification of histone proteins by methylation plays a central role in chromatin regulation by recruiting epigenetic "readers" via specialized binding domains. Depending on the degree of methylation, the exact modified amino acid, and the associated reader proteins histone methylations are involved in the regulation of all DNA-based processes, such as transcription, DNA replication, and DNA repair. Here we present methods to identify histone methylation readers using a mass spectrometry-linked nucleosome affinity purification approach. We provide detailed protocols for the generation of semisynthetic methylated histones, their assembly into biotinylated nucleosomes, and the identification of methylation-specific nucleosome-interacting proteins from nuclear extracts via nucleosome pull-downs and label-free quantitative proteomics. Due to their versatility, these protocols allow the identification of readers of various histone methylations, and can also be adapted to different cell types and tissues, and other types of modifications.
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Publikationstyp Artikel: Sammelbandbeitrag/Buchkapitel
Korrespondenzautor
Schlagwörter Affinity Purification ; Chromatin ; Histone ; Histone Modification ; Mass Spectrometry ; Methylation ; Native Chemical Ligation ; Nuclear Extract ; Nucleosome ; Proteomics
ISSN (print) / ISBN 1064-3745
e-ISSN 1940-6029
Bandtitel Histone Methyltransferases
Zeitschrift Methods in Molecular Biology
Quellenangaben Band: 2529, Heft: , Seiten: 327-403 Artikelnummer: , Supplement: ,
Verlag Springer
Verlagsort Berlin [u.a.]
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Functional Epigenetics (IFE)