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):

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