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Characterization of Complete Histone Tail Proteoforms Using Differential Ion Mobility Spectrometry
Pavel V Shliaha1, Matthew A Baird2, Mogens M Nielsen1
1Department of Biochemistry and Molecular Biology, VILLUM Center for Bioanalytical Sciences, and Center for Epigenetics, University of Southern Denmark , DK-5230 Odense M, Denmark.
Analytical Chemistry
|April 14, 2017
Summary
Researchers used high-definition differential ion mobility spectrometry to separate and identify histone tails with various post-translational modifications (PTMs). This method successfully resolved large variant peptides, advancing proteomics research.
Area of Science:
- Biochemistry
- Proteomics
- Chromatin Biology
Background:
- Histone proteins undergo dynamic post-translational modifications (PTMs) that regulate chromatin structure and function.
- Functional PTMs are primarily located on the N-terminal histone tails, approximately 50-residue domains extending from the nucleosome core.
Purpose of the Study:
- To develop a method for rapid gas-phase separation and identification of histone tails with various PTMs.
- To demonstrate the capability of resolving large variant peptides with subtle mass differences due to PTMs.
Main Methods:
- Utilized high-definition differential ion mobility spectrometry (HD-IMS) coupled with electron transfer dissociation (ETD).
- Analyzed histone tails with monomethylation, trimethylation, acetylation, and phosphorylation.
Main Results:
- Achieved rapid baseline gas-phase separation of histone tails with distinct PTMs.
- Successfully identified variant peptides, including those where PTMs contributed as little as 0.25% to the total mass.
- Resolved the largest variant peptides reported to date using this technique.
Conclusions:
- HD-IMS with ETD provides a powerful approach for analyzing complex histone PTMs.
- This method has significant implications for top-down proteomics and the analysis of intact proteins with PTMs.