分子转子作为条件光标签,用于快速检测生物分子相互作用
Walter L Goh1, Min Yen Lee, Thomas L Joseph
1Molecular Engineering Laboratory, Biomedical Sciences Institutes, A*STAR , 61 Biopolis Drive, #03-12 Proteos, Singapore 138673.
Journal of the American Chemical Society
|February 6, 2014
概括
光分子转子现在可以检测特定的蛋白相互作用. 一个新的p53记者 (JP1-R) 在Mdm2结合时有条件光,从而能够有效地选相互作用抑制剂.
科学领域:
- 生物化学 生物化学
- 化学生物学 化学生物学
- 分子生物学分子生物学
背景情况:
- 光分子转子通常用作粘度探针.
- 它们在检测特定生物分子相互作用中的应用仍然未被探索.
- p53-Mdm2相互作用是研究-蛋白结合的一个特征很好的模型系统.
研究的目的:
- 探索光分子旋转器用于检测特定-蛋白相互作用的实用性.
- 为p53-Mdm2相互作用开发一种新的光记者.
- 为选p53-Mdm2相互作用抑制剂建立一个均的试验法.
主要方法:
- 基于朱洛利丁的p53记者 (JP1-R) 的设计和合成.
- 在同质测试中使用JP1-R来选一个片段库.
- 使用已建立的二次测定验证已识别的抑制剂.
主要成果:
- JP1-R 呈现条件光,仅在与 Mdm2.2 结合时发射.
- 使用JP1-R的片段图书馆屏幕识别了p53-Mdm2相互作用的几个对手.
- 与已建立的二次试验相比,JP1-R试验表现出更高的灵敏度.
结论:
- 光分子转子是用于检测特定生物分子相互作用的多功能探针.
- JP1-R是识别p53-Mdm2相互作用的抑制剂的有效工具.
- 这种方法为生物分子相互作用研究提供了一种敏感和均的方法.
相关概念视频
01:19Protein Dynamics in Living Cells
1.9K
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...
1.9K
03:31Labeling DNA Probes
7.7K
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
7.7K