Profiling Changes in Histone Post-translational Modifications by Top-Down Mass Spectrometry
Mowei Zhou1, Si Wu1,2, David L Stenoien1
1Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|November 11, 2016
Summary
Top-down mass spectrometry (MS) rapidly profiles histone proteoforms and their modifications. This method enables comparative analysis of gene expression changes between different biological samples, like wild-type versus mutant species.
Area of Science:
- Biochemistry
- Genomics
- Proteomics
Background:
- Gene expression is regulated by histone post-translational modifications, collectively known as the histone code.
- Understanding the histone code is crucial for deciphering cellular functions and disease mechanisms.
Purpose of the Study:
- To present a top-down mass spectrometry (MS) workflow for rapid, global profiling of histone proteoforms.
- To demonstrate the utility of this workflow for comparative analysis of histone modifications between different biological states.
Main Methods:
- A top-down mass spectrometry workflow utilizing liquid chromatography (LC) coupled to MS (LC-MS) was developed.
- The method was applied to compare histone proteoforms between a wild-type and a mutant fungal species.
Main Results:
- The LC-MS top-down approach enabled fast global profiling of histone proteoforms.
- Differential abundances of specific proteoforms were identified between the wild-type and mutant fungal species.
Conclusions:
- This top-down LC-MS method provides a powerful tool for analyzing the histone code and identifying changes in histone modifications.
- The protocol is adaptable for screening histone modification alterations in various comparative studies, including wild-type vs. mutant or healthy vs. diseased samples.
Keywords:
HistoneLiquid chromatographyMass spectrometryPost-translational modificationScreeningTop-down
