Application of histone modification-specific interaction domains as an alternative to antibodies
- Goran Kungulovski1,
- Ina Kycia1,4,
- Raluca Tamas1,
- Renata Z. Jurkowska1,
- Srikanth Kudithipudi1,
- Chisato Henry2,
- Richard Reinhardt3,
- Paul Labhart2 and
- Albert Jeltsch1
- 1Institute of Biochemistry, Stuttgart University, 70569 Stuttgart, Germany;
- 2Active Motif, Carlsbad, California 92008, USA;
- 3Max-Planck-Genomzentrum Köln, 50829 Köln, Germany
- Corresponding author: albert.jeltsch{at}ibc.uni-stuttgart.de
Abstract
Post-translational modifications (PTMs) of histones constitute a major chromatin indexing mechanism, and their proper characterization is of highest biological importance. So far, PTM-specific antibodies have been the standard reagent for studying histone PTMs despite caveats such as lot-to-lot variability of specificity and binding affinity. Herein, we successfully employed naturally occurring and engineered histone modification interacting domains for detection and identification of histone PTMs and ChIP-like enrichment of different types of chromatin. Our results demonstrate that histone interacting domains are robust and highly specific reagents that can replace or complement histone modification antibodies. These domains can be produced recombinantly in Escherichia coli at low cost and constant quality. Protein design of reading domains allows for generation of novel specificities, addition of affinity tags, and preparation of PTM binding pocket variants as matching negative controls, which is not possible with antibodies.
Footnotes
-
[Supplemental material is available for this article.]
-
Article published online before print. Article, supplemental material, and publication date are at http://www.genome.org/cgi/doi/10.1101/gr.170985.113.
- Received December 11, 2013.
- Accepted August 7, 2014.
This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genome.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
