PXD048061

PXD048061 is an original dataset announced via ProteomeXchange.

Title	Coupling SDS-PAGE to capillary zone electrophoresis-tandem mass spectrometry for high-resolution top-down proteomics analysis of intact histone proteoforms
Description	Top-down mass spectrometry (TDMS) is a powerful tool to reveal the mechanism of epigenetic regulation by histones. However, the high similarity of histone variants and their highly dynamic post-translational modifications (PTMs) make them as on-going challenge analytes for separation with traditional chromatographic-based methods. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE), which enables high-resolution protein separation based on molecular size, not only split histone proteins in different channels, but also isolate the large intact histone proteins from the truncated ones. Here, for the first time, we employed SDS-PAGE integrating with the second-dimensional separation of capillary zone electrophoresis (CZE) to distinguish intact histone proteoforms containing different PTMs. Due to the unique physicochemical property of histone proteins regarding small proteins carrying highly positive charges, we systematically evaluated every step in the strategy, achieving high histone protein recovery with trichloroacetic acid precipitation, better separation with basic sample buffer as well as confident identification with low collision energy for histone proteins. The optimized workflow provides the distinguish of intact histone variants that differ by few amino acids near both the N- and C-termini as well as the identification of combinatorial PTMs and the crosstalk between PTMs.
HostingRepository	PRIDE
AnnounceDate	2024-10-15
AnnouncementXML	Submission_2024-10-15_05:25:51.229.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Liangliang Sun
SpeciesList	scientific name: Bos taurus (Bovine); NCBI TaxID: 9913;
ModificationList	phosphorylated residue; acetylated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2023-12-21 18:19:36	ID requested
⏵ 1	2024-10-15 05:26:01	announced

10.1002/pmic.202300650;

Fang F, Gao G, Wang Q, Wang Q, Sun L, Combining SDS-PAGE to capillary zone electrophoresis-tandem mass spectrometry for high-resolution top-down proteomics analysis of intact histone proteoforms. Proteomics, 24(17):e2300650(2024) [pubmed]

submitter keyword: SDS-PAGE, top-down proteomics, post-translational modifications, histone proteoform,CZE-MS/MS

Liangliang Sun
contact affiliation	Department of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI 48824, United States.
contact email	lsun@chemistry.msu.edu
lab head
Liangliang Sun
contact affiliation	Michigan State University
contact email	lsun@chemistry.msu.edu
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD048061
     1. Label: PRIDE project
     2. Name: Coupling SDS-PAGE to capillary zone electrophoresis-tandem mass spectrometry for high-resolution top-down proteomics analysis of intact histone proteoforms