细胞内光催化接近标记(iPPL)用于靶向组蛋白H3的染色质结合蛋白的动态分析

IF 3.5 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Kazuki Miura, Hikaru Niimi, Tatsuya Niwa, Hideki Taguchi and Hiroyuki Nakamura*, 
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引用次数: 0

摘要

我们展示了一种利用细胞内光催化接近标记(iPPL)分析组蛋白H3蛋白-蛋白相互作用(PPI)的新方法。结果表明,以吖啶黄为光催化剂,以1-甲基-4-芳基脲唑(MAUra)为蛋白质标记剂的组合是进行蛋白质接近标记反应的最有效策略。此外,对组蛋白H3相互作用蛋白组蛋白赖氨酸n -甲基转移酶EZH2中标记氨基酸的鉴定表明,EZH2中距离组蛋白H3几纳米范围内的氨基酸被iPPL标记。与传统的接近标记方法相比,这种受限的标记半径允许更集中的PPI分析。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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

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

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.

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来源期刊
ACS Chemical Biology
ACS Chemical Biology 生物-生化与分子生物学
CiteScore
7.50
自引率
5.00%
发文量
353
审稿时长
3.3 months
期刊介绍: ACS Chemical Biology provides an international forum for the rapid communication of research that broadly embraces the interface between chemistry and biology. The journal also serves as a forum to facilitate the communication between biologists and chemists that will translate into new research opportunities and discoveries. Results will be published in which molecular reasoning has been used to probe questions through in vitro investigations, cell biological methods, or organismic studies. We welcome mechanistic studies on proteins, nucleic acids, sugars, lipids, and nonbiological polymers. The journal serves a large scientific community, exploring cellular function from both chemical and biological perspectives. It is understood that submitted work is based upon original results and has not been published previously.
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