Dataset Information

Site-Link for identification of chromatinized proteome defined by site-specific histone acylations in living cells

ABSTRACT: By coupling the mass spectrometry, we developed the Site-Link strategy based on the GCE method for analyzing the interacting proteome of histone lysine acylations in living cells. As K* could mimic the natural lysine to be recognized by natural lysine synthase, histones bearing the site-specifically incorporated K*acyl, should also be also integrated into nucleosomes and further compacted into chromatin similar as Kacyl with cell passaging. The subsequent 365-nm light irradiation could activate the diazirine and covalently capture potential effector proteins on native chromatin in living cells. The effector proteins could be identified after gel-based proteomics and MS/MS analysis of the crosslinked histone-effector complexes.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Early Embryonic Cell

SUBMITTER: Fangfei Qin

LAB HEAD: Peng R. Chen

PROVIDER: PXD033959 | Pride | 2023-03-06

REPOSITORIES: pride

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Publications

Linking chromatin acylation mark-defined proteome and genome in living cells.

Qin Fangfei F Li Boyuan B Wang Hui H Ma Sihui S Li Jiaofeng J Liu Shanglin S Kong Linghao L Zheng Huangtao H Zhu Rongfeng R Han Yu Y Yang Mingdong M Li Kai K Ji Xiong X Chen Peng R PR

Cell 20230301 5

A generalizable strategy with programmable site specificity for in situ profiling of histone modifications on unperturbed chromatin remains highly desirable but challenging. We herein developed a single-site-resolved multi-omics (SiTomics) strategy for systematic mapping of dynamic modifications and subsequent profiling of chromatinized proteome and genome defined by specific chromatin acylations in living cells. By leveraging the genetic code expansion strategy, our SiTomics toolkit revealed di ...[more]

PMID: 36868209

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