Proteomics

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Middle-Down Characterization of the Cell Cycle Dependence of Histone H4 PTMs and Proteoforms


ABSTRACT: In the present study, a middle-down approach is employed for the identification, characterization, and quantitation of the PTMs and proteoforms of histone H4 in two breast cancer cell lines. Our experimental strategy not only preserves the combinatorial attributes of existing PTMs, but also enhances the separation ability and reduces the complexity of downstream data analysis. The fluctuations in the relative abundances of common modifications throughout the cell cycle, including phosphorylation, acetylation, and methylation, suggest that they play significant roles at different stages of the cell cycle in breast cancer and could participate in carcinogenesis. In summary, our analysis advances the understanding and elucidation of cancer epigenetics at the single-molecule level, and lays the foundation for further study of histones or other proteins.

INSTRUMENT(S): Fourier Transform Ion Cyclotron Resonance Mass Spectrometer

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

DISEASE(S): Breast Cancer

SUBMITTER: Tingting Jiang  

LAB HEAD: Nicolas L. Young

PROVIDER: PXD008296 | Pride | 2018-07-05

REPOSITORIES: Pride

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Publications

Middle-Down Characterization of the Cell Cycle Dependence of Histone H4 Posttranslational Modifications and Proteoforms.

Jiang Tingting T   Hoover Michael E ME   Holt Matthew V MV   Freitas Michael A MA   Marshall Alan G AG   Young Nicolas L NL  

Proteomics 20180601 11


Post-translational modifications (PTMs) of histones are important epigenetic regulatory mechanisms that are often dysregulated in cancer. We employ middle-down proteomics to investigate the PTMs and proteoforms of histone H4 during cell cycle progression. We use pH gradient weak cation exchange-hydrophilic interaction liquid chromatography (WCX-HILIC) for on-line liquid chromatography-mass spectrometry analysis to separate and analyze the proteoforms of histone H4. This procedure provides enhanc  ...[more]

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