可点击的光反应性ATP亲和探头用于ATP结合蛋白的全球分析
Zhiming Wang1,2, Jing Tan1,2, Mengxuan Li1,3
1Institute of Pharmaceutical Analysis, Key Laboratory of Chemical Biology (MOE), School of Pharmaceutical Sciences, Cheeloo College of Medicine, Shandong University, Jinan 250012, Shandong, China.
Analytical chemistry
|November 22, 2023
概括
研究人员开发了一种可点击的ATP光 afinity探针,以识别腺三酸盐 (ATP) 结合蛋白. 该工具有助于为疾病研究分析ATP结合蛋白和激酶抑制剂标.
科学领域:
- 生物化学 生化学
- 化学生物学 化学生物学
- 蛋白质组学是指蛋白质组学.
背景情况:
- 腺三酸盐 (ATP) 对于细胞能量转移至关重要.
- 激酶,ATP结合蛋白,调节细胞信号,并与癌症等疾病有关.
- 选择性激酶抑制剂是重要的治疗方法,但非目标效应需要确定目标.
研究的目的:
- 开发一个可点击的ATP光亲和探针,用于ATP结合蛋白的全球分析.
- 建立一种化学蛋白质学方法,用于对酶抑制剂的高通量标识.
主要方法:
- 用可点击的ATP光亲和探针化蛋白质溶解酸.
- 紫外线照射以标记ATP结合蛋白与基因手柄.
- 生物素结合通过点击化学,斯特雷普塔维丁丰富,素消化和LC-MS/MS分析.
主要成果:
- 在单个LC-MS/MS运行中识别了400多种ATP结合蛋白,包括大约200种激酶.
- 通过无标签量化,成功识别了三种激酶抑制剂的报告目标.
- 通过Western blot分析验证识别的目标.
结论:
- 一个可点击的ATP光亲和探针使ATP结合蛋白的蛋白质全基因定型成为可能.
- 这种化学蛋白质组方法可促进对酶抑制剂的高通量标识.
- 这种方法对于理解酶信号通路和开发向疗法非常有价值.
相关概念视频
03:31Labeling DNA Probes
8.2K
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...
8.2K
01:19Protein Dynamics in Living Cells
2.1K
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...
2.1K