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Preparative two-step purification of recombinant H1.0 linker histone and its domains

Ivić, Nives; Bilokapić, Silvija; Halić, Mario (2017) Preparative two-step purification of recombinant H1.0 linker histone and its domains. PLoS One, 12 (12). ISSN 1932-6203

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Abstract

H1 linker histones are small basic proteins that have a key role in the formation and maintenance of higher-order chromatin structures. Additionally, many examples have shown that linker histones play an important role in gene regulation, modulated by their various subtypes and posttranslational modifications. Obtaining high amounts of very pure linker histones, especially for efficient antibody production, remains a demanding and challenging procedure. Here we present an easy and fast method to purify human linker histone H1.0 overexpressed in Escherichia coli, as well as its domains: N-terminal/globular domain and C-terminal intrinsically disordered domain. This purification protocol relies on a simple affinity chromatography step followed by cation exchange due to the highly basic properties of histone proteins. Therefore, this protocol can also be applied to other linker histones. Highly pure proteins in amounts sufficient for most biochemical experiments can be obtained. The functional quality of purified H1.0 histone and its domains has been confirmed by pull-down, gel-mobility shift assays and the nuclear import assay.

Item Type: Article
Uncontrolled Keywords: linker histone, purification, nucleosome, chromatin
Subjects: NATURAL SCIENCES > Biology
Divisions: Division of Molecular Biology
Division of Physical Chemistry
Projects:
Project titleProject leaderProject codeProject type
291823 Marie Curie FP7-PEOPLE-2011-COFUNDUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Depositing User: Nives Ivić
Date Deposited: 19 Jan 2018 09:41
Last Modified: 19 Jan 2018 09:41
URI: http://fulir.irb.hr/id/eprint/3843
DOI: 10.1371/journal.pone.0189040.

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