Introduction

As histones play central roles in most chromosomal functions including regulation of DNA replication, DNA damage repair, and gene transcription, both their basic biology and their roles in disease development have been the subject of intense study. Because multiple post-translational modifications (PTMs) along the entire protein sequence are potential regulators of histones, a top-down approach, where intact proteins are analyzed, is ultimately required for complete characterization of proteoforms. However, significant challenges remain for top-down histone analysis primarily because of deficiencies in separation/resolving power and effective identification algorithms. Here we used state-of-the-art mass spectrometry and a bioinformatics workflow for targeted data analysis and visualization. The workflow uses ProMex for intact mass deconvolution, MSPathFinder as a search engine, and LcMsSpectator as a data visualization tool. When complemented with the open-modification tool TopPIC, this workflow enabled identification of novel histone PTMs including tyrosine bromination on histone H4 and H2A, H3 glutathionylation, and mapping of conventional PTMs along the entire protein for many histone subunits.

Publications

  1. Profiling of Histone Post-Translational Modifications in Mouse Brain with High-Resolution Top-Down Mass Spectrometry.
    Cite this
    Zhou M, Paša-Tolić L, Stenoien DL, 2016-12-01 - Journal of proteome research

Credits

  1. Mowei Zhou
    Developer

    Pacific Northwest National Laboratory, Earth and Biological Sciences Directorate, United States of America

  2. Ljiljana Paša-Tolić
    Developer

    Pacific Northwest National Laboratory, Earth and Biological Sciences Directorate, United States of America

  3. David L Stenoien
    Investigator

    Pacific Northwest National Laboratory, Earth and Biological Sciences Directorate, United States of America

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Summary
AccessionBT000441
Tool TypeApplication
Category
PlatformsLinux/Unix
TechnologiesPerl
User InterfaceTerminal Command Line
Download Count0
Country/RegionUnited States of America
Submitted ByDavid L Stenoien