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Enhanced top-down characterization of histone post-translational modifications |
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| Authors: | Zhixin Tian Nikola Toli? Rui Zhao Ronald J Moore Shawna M Hengel Errol W Robinson David L Stenoien Si Wu Richard D Smith Ljiljana Pa?a-Toli? |
| Institution: | 1Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, 902 Battelle Blvd, Richland, WA 99352, USA;2Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 457 Zhongshan Road, Dalian, Liaoning 116023, China;3Biological Sciences Division, Pacific Northwest National Laboratory, 902 Battelle Blvd, Richland, WA 99352, USA |
| 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 |