Identification and profiling of dynamic changes of site-specific histone acylations in HEK293T cells under metabolites treatment

ABSTRACT: 1. Systematic survey of PTM site and stoichiometry on histone proteins has been lagged behind due to the lack of efficient quantitative peptide comparison methodology on histone proteins. Our quantitative mass spectrometry-based proteomics approach, Site-Profiling, used SILAC-based peptide ratio analysis on histone acylations with and without SCFA-derived metabolic treatment to directly compare the same peptide with the same modification and the same chain length. Different from the published methods that isotopically labeled metabolites, which might cause false-positive results and information lost, we only labelled all the lysines and arginines in histones and thus the same modification under native conditions would be kept, resulting that all modification peptides were comparable in parallel before and after metabolites stimulation. By a comprehensive analysis of all peptides containing the same histone marks, Site-Map can compare the abundance of each site-specific modification upon the treatment of the corresponding metabolite. 2. To reveal the site dependency of histone Kbhb distributed at the super enhancer region, we developed a method to isolate the super enhancer condensates by co-immunoprecipitation with coactivators BRD4. Same site-mapping method was utilized for comparasion the site dependency of histone Kbhb enriched with BRD4 or not.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Early Embryonic Cell

DISEASE(S): Disease Free

SUBMITTER: Fangfei Qin  

LAB HEAD: Peng R. Chen

PROVIDER: PXD029508 | Pride | 2023-03-06

REPOSITORIES: Pride