Application of histone modification-specific interaction domains as an alternative to antibodies

Kungulovski, Goran, Kycia, Ina, Tamas, Raluca, Jurkowska, Renata Z. ORCID: https://orcid.org/0000-0002-4507-2222, Kudithipudi, Srikanth, Henry, Chisato, Reinhardt, Richard, Labhart, Paul and Jeltsch, Albert 2014. Application of histone modification-specific interaction domains as an alternative to antibodies. Genome Research 24 (11) , pp. 1842-1853. 10.1101/gr.170985.113

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Abstract

Post-translational modifications (PTMs) of histones constitute a major chromatin indexing mechanism, and their proper characterizationis of highest biological importance. So far, PTM-specific antibodies have been the standard reagent for studying histone PTMsdespite caveats such as lot-to-lot variability of specificity and binding affinity. Herein, we successfully employed naturallyoccurring and engineered histone modification interacting domains for detection and identification of histone PTMs and ChIP-likeenrichment of different types of chromatin. Our results demonstrate that histone interacting domains are robust and highlyspecific reagents that can replace or complement histone modification antibodies. These domains can be produced recombinantlyin Escherichia coli at low cost and constant quality. Protein design of reading domains allows for generation of novel specificities, additionof affinity tags, and preparation of PTM binding pocket variants as matching negative controls, which is not possible withantibodies.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Biosciences
Additional Information:	Available under a Creative Commons License (Attribution-NonCommercial 4.0 International).
Publisher:	Cold Spring Harbor Laboratory Press
ISSN:	1088-9051
Date of First Compliant Deposit:	31 October 2019
Date of Acceptance:	7 August 2014
Last Modified:	05 May 2023 09:28
URI:	https://orca.cardiff.ac.uk/id/eprint/126454

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