在活细胞和生物体中使用非扩散性近距离标记系统识别目标蛋白质
Yingjie Sun1, Changheng Li1, Xiaofei Deng2
1Department of Neurology, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, China.
eLife
|December 27, 2024
概括
我们开发了POST-IT,这是一种新的活细胞近距离标记系统,用于在它们的原生细胞环境中识别药物点. 这项技术成功地发现了新的药物向相互作用,推动了小分子药物发现.
科学领域:
- 生物化学 生物化学
- 化学生物学 化学生物学
- 药物发现 药物发现 药物发现
背景情况:
- 准确识别药物标蛋白对于药物开发和理解药物机制至关重要.
- 现有的标识别方法往往无法捕捉完整的细胞环境中的相互作用,从而限制了像短暂复合体和膜蛋白等标的发现.
研究的目的:
- 开发一种新的近距离标记系统,用于在活细胞中识别小分子目标.
- 通过保持原生细胞环境来克服基于细胞解质的方法的局限性.
主要方法:
- 开发了POST-IT (Pup-On-target for Small molecule Target Identification Technology),一种非扩散性近距离标记系统,使用工程保护体辅助因子A和HaloTag.
- 优化了系统,以防止自我毛皮化,多毛皮化和脱毛,并改进了化学链接器.
- 在活细胞和活斑马鱼胚胎中应用POST-IT.
主要成果:
- POST-IT成功地确定了已知的毒品目标.
- 发现SEPHS2是达沙替尼的新目标,VPS37C是氧化的新目标.
- 更好地了解这些药物的作用机制.
结论:
- POST-IT是一种有效的技术,用于在活细胞环境中识别小分子标.
- 该系统对生物研究和推进药物发现具有广泛的潜力,正如斑马鱼胚胎中所示的那样.
关键词:
现在,我们可以说PafA.生物化学 生物化学化学生物学 化学生物学克洛洛奎因 (Chloroquine) 是一种药物达萨提尼布 (dasatinib) 是一种接近性标签的标签.目标识别 目标识别斑马鱼是一种斑马鱼.更多相关视频
11:16Discovering Protein Interactions and Characterizing Protein Function Using HaloTag Technology
Published on: July 12, 2014
36.9K
11:04Inducible LAP-tagged Stable Cell Lines for Investigating Protein Function, Spatiotemporal Localization and Protein Interaction Networks
Published on: December 24, 2016
9.8K
相关概念视频
01:19Protein Dynamics in Living Cells
2.0K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.0K
01:24Tagging and Fusion Proteins
6.5K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
6.5K