Combining SDS-PAGE with Capillary Zone Electrophoresis-Tandem Mass Spectrometry for Top-Down Proteomics Analysis of Intact Histone Proteoforms.

Authors

Fei Fang, Liangliang Sun

Published in

Methods in molecular biology (Clifton, N.J.). Volume 3018. Pages 185-198.

Abstract

Mass spectrometry (MS)-based top-down proteomics (TDP) analysis of histone proteoforms provides critical information about combinatorial posttranslational modifications (PTMs), which is vital for pursuing a better understanding of epigenetic regulation of gene expression. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) separates histone proteins (H1, H2, H3, and H4) based on their molecular weight, while capillary zone electrophoresis (CZE) provides additional separation of proteoforms of each histone protein based on their electrophoretic mobility, which is affected by PTMs, e.g., acetylation and phosphorylation. Here, we describe the combination of SDS-PAGE-based protein fractionation with CZE-tandem MS (MS/MS) for high-resolution characterization of histone proteoforms. Over 200 histone proteoforms from a commercial calf thymus histone sample were identified with good reproducibility.

PMID:
42542530
Bibliographic data and abstract were imported from PubMed on 02 Aug 2026.

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