相关实验视频

Updated: Mar 9, 2026

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein

Published on: June 30, 2019

在活细胞中使用素化探针进行广谱激酶分析

Qian Zhao, Xiaohu Ouyang, Xiaobo Wan

¹Worldwide Research and Development, Pfizer , San Diego, California 92121, United States.

Journal of the American Chemical Society

|January 5, 2017

概括

科学家们开发了新的硫化探针来标记活细胞中的许多蛋白质激酶. 这些探针能够精确地测量药物与激酶的相互作用,从而推动化学蛋白质组学研究.

科学领域:

生物化学
化学生物学
药理学

背景情况:

蛋白激酶是关键的药物点, 但选择性抑制是具有挑战性的.
在活细胞中量化药物相互作用是很困难的.

研究的目的:

设计和验证用于广泛基因组标记的硫化物探针.
使用这些探针量化药物对细胞内激酶的作用.

主要方法:

酸硫探针的设计,以准化在酶ATP结合位点中的保存化残留物.
蛋白质结晶学和质谱学以确认探测器的反应性.
应用优化探针 (XO44) 和无标记质谱来研究达沙替尼的作用.

主要成果:

硫化探针共价标记广泛的内源性激酶.
优化探针2 (XO44) 修改了133个激酶,与细胞内ATP竞争.
在相关度下精确量化达沙替尼与特定酶的结合.

结论:

素化探针是化学蛋白质应用的有效工具.
这种方法可以精确地测量活细胞中的药物向作用.
有助于理解激酶抑制剂的选择性和有效性.

更多相关视频

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

Monitoring Kinase and Phosphatase Activities Through the Cell Cycle by Ratiometric FRET

Published on: January 27, 2012

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Identification of Novel CK2 Kinase Substrates Using a Versatile Biochemical Approach

Published on: February 21, 2019

相关概念视频

Labeling DNA Probes

Labeling DNA Probes

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...