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

Autor: Miura K; Laboratory for Chemistry and Life Science, Institute of Integrated Research, Institute of Science Tokyo, Yokohama 226-8501, Japan.; School of Life Science and Technology, Institute of Science Tokyo, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan., Niimi H; School of Life Science and Technology, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan., Niwa T; School of Life Science and Technology, Institute of Science Tokyo, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.; Cell Biology Center, Institute of Integrated Research, Institute of Science Tokyo, 4259, Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan., Taguchi H; School of Life Science and Technology, Institute of Science Tokyo, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.; Cell Biology Center, Institute of Integrated Research, Institute of Science Tokyo, 4259, Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan., Nakamura H; Laboratory for Chemistry and Life Science, Institute of Integrated Research, Institute of Science Tokyo, Yokohama 226-8501, Japan.; School of Life Science and Technology, Institute of Science Tokyo, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Jazyk: angličtina
Zdroj: ACS chemical biology [ACS Chem Biol] 2024 Dec 20; Vol. 19 (12), pp. 2412-2417. Date of Electronic Publication: 2024 Dec 09.
DOI: 10.1021/acschembio.4c00680
Abstrakt: 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.
Databáze: MEDLINE
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