This article is part of a special issue on epigenomics.

Open Access Method

Enhanced top-down characterization of histone post-translational modifications

Zhixin Tian12, Nikola Tolić1, Rui Zhao1, Ronald J Moore3, Shawna M Hengel1, Errol W Robinson1, David L Stenoien3, Si Wu1, Richard D Smith3 and Ljiljana Paša-Tolić1*

Author affiliations

1 Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, 902 Battelle Blvd, Richland, WA 99352, USA

2 Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, Liaoning 116023, China

3 Biological Sciences Division, Pacific Northwest National Laboratory, 902 Battelle Blvd, Richland, WA 99352, USA

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Citation and License

Genome Biology 2012, 13:R86  doi:10.1186/gb-2012-13-10-r86

Published: 3 October 2012

Abstract

Post-translational modifications (PTMs) of core histones work synergistically to fine tune chromatin structure and function, generating a so-called histone code that can be interpreted by a variety of chromatin interacting proteins. We report a novel online two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS/MS) platform for high-throughput and sensitive characterization of histone PTMs at the intact protein level. The platform enables unambiguous identification of 708 histone isoforms from a single 2D LC-MS/MS analysis of 7.5 µg purified core histones. The throughput and sensitivity of comprehensive histone modification characterization is dramatically improved compared with more traditional platforms.

Keywords:
Saltless WCX-HILIC; top-down; histone; posttranslational modification