Middle-down proteomic analyses with Ion mobility separations of endogenous isomeric proteoforms

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Authors
Shliaha, Pavel V.
Gorshkov, Vladimir A.
Kovalchuk, Sergey I.
Schwammle, Veit
Baird, Matthew A.
Shvartsburg, Alexandre A.
Jensen, Ole N.
Advisors
Issue Date
2020-01-14
Type
Article
Keywords
Mass spectrometry , Peptides and proteins , ChromatographyIons , Molecular structure
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Citation
Shliaha, Pavel V.; Gorshkov, Vladimir A.; Kovalchuk, Sergey I.; Schwammle, Veit; Baird, Matthew A.; Shvartsburg, Alexandre A.; Jensen, Ole N. 2020. Middle-down proteomic analyses with Ion mobility separations of endogenous isomeric proteoforms. Analytical chemistry, vol. 92:no. 3:pp 2364-2368
Abstract

Biological functions of many proteins are governed by post-translational modifications (PTMs). In particular, the rich PTM complement in histones controls the gene expression and chromatin structure with major health implications via a combinatoric language. Deciphering that "histone code" is the great challenge for proteomics given an astounding number of possible proteoforms, including isomers with different PTM positions. These must be disentangled on the top- or middle-down level to preserve the key PTM connectivity, which condensed-phase separations failed to achieve. We reported the capability of ion mobility spectrometry (IMS) methods to resolve such isomers for model histone tails. Here, we advance to biological samples, showing middle-down analyses of histones from mouse embryonic stem cells via online chromatography to fractionate proteoforms with distinct PTM sets, differential or field asymmetric waveform IMS (FAIMS) to resolve the isomers, and Orbitrap mass spectrometry with electron transfer dissociation to identify the resolved species.

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Publisher
NLM (Medline)
Journal
Book Title
Series
Analytical chemistry;v.92:no.3
PubMed ID
DOI
ISSN
1520-6882
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