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E-raamat: Histone Methyltransferases: Methods and Protocols

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This volume provides methods used to investigate histone methyltransferase function. Chapters guide readers through a comprehensive set of approaches that detail phylogenetic diversity, histone demethylase activities in vitro, generating chromatin substrates, auto-methylation, quantification of metabolites, protein purification, crystallization, X-ray structure, cryogenic electron microscopy, assessing genome-wide patterns, CUT&Tag in mouse embryonic tissues, chemical biology approaches, peptide SPOT arrays, nascent chromatin capture, ectopic protein tethering, computational models, and development of methyltransferase inhibitors. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.





 





Authoritative and cutting-edge, Histone Methyltransferases: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
Not all is SET for methylation: Evolution of eukaryotic protein
methyltransferases.- Detection and quantification of histone
methyltransferase activity in vitro.-  In Vitro Histone Demethylase
Assays.- Preparation and characterization of chromatin templates for histone
methylation assays.- Techniques to study automethylation of histone
methyltransferases and its functional impact.- Rodriguez Profiling the
regulation of histone methylation and demethylation by metabolites and
metals.- Determination of histone methyltransferase structure by
crystallography.- Determination of histone methyltransferase structures in
complex with the nucleosome by cryogenic electron microscopy.- Development
and validation of antibodies targeting site-specific histone
methylation.- Genetic, genomic and imaging approaches to dissect the role of
Polycomb group epigenetic regulators in mice.- Profiling histone methylation
in low numbers of cells.-  Automated CUT&RUN using the KingFisher Duo
Prime.- Bioinformatics methods for ChIP-seq histone
analysis.- Characterization of SET-domain histone lysine methyltransferase
substrates using a cofactor S-adenosyl-L-methionine surrogate.- Specificity
analysis of protein methyltransferases and discovery of novel substrates
using SPOT peptide arrays.- Identifying specific protein interactors of
nucleosomes carrying methylated histones using mass
spectrometr.- Investigating mitotic inheritance of histone post-translational
modifications by triple pSILAC coupled to Nascent Chromatin
Capture.- Investigating mitotic inheritance of histone modifications using
tethering.- Investigating histone modification dynamics by mechanistic
computational modelling.- Screening for small molecule inhibitors of histone
methyltransferases.
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