Intracellular Photocatalytic Proximity Labeling (iPPL) for Dynamic Analysis of Chromatin-Binding Proteins Targeting Histone H3

ACS Chem Biol. 2024 Dec 20;19(12):2412-2417. doi: 10.1021/acschembio.4c00680. Epub 2024 Dec 9.

Abstract

We demonstrated a novel approach for protein-protein interaction (PPI) profiling of histone H3 using intracellular photocatalytic-proximity labeling (iPPL). This approach identified that the combination of acriflavine as a photocatalyst and 1-methyl-4-arylurazol (MAUra) as a protein labeling agent was the most efficient strategy to proceed the protein proximity labeling reaction. Furthermore, the identification of the labeled amino acids in histone H3 interacting proteins, histone lysine N-methyltransferase EZH2, showed that the amino acid in EZH2 within a few nanometers from histone H3 is labeled by iPPL. This restricted labeling radius allows for more-focused PPI profiling, compared to conventional proximity labeling methods.

MeSH terms

  • Catalysis
  • Chromatin / chemistry
  • Chromatin / metabolism
  • Enhancer of Zeste Homolog 2 Protein / metabolism
  • Histones* / chemistry
  • Histones* / metabolism
  • Humans
  • Photochemical Processes
  • Protein Binding
  • Staining and Labeling / methods

Substances

  • Histones
  • Chromatin
  • Enhancer of Zeste Homolog 2 Protein
  • EZH2 protein, human