Department of Chemistry, School of Physics and Chemistry, Gwangju Institute of Science and Technology, Gwangju 61005, Republic of Korea.
Department of Global Medical Science, Sungshin University, Seoul 01133, Republic of Korea.
ACS Chem Biol. 2020 Sep 18;15(9):2346-2354. doi: 10.1021/acschembio.0c00279. Epub 2020 Aug 19.
The high temperature requirement A (HTRA) family of serine proteases mediates protein quality control. These proteins process misfolded proteins in several diseases including Alzheimer's disease (AD) and Parkinson's disease (PD). While their structures and activation mechanisms have been studied, the precise details of the regulation of their activity under physiological conditions have not been completely elucidated, partly due to the lack of suitable chemical probes. In the present study, we developed novel activity-based probes (ABPs) targeting the HTRAs and demonstrated their utility in the monitoring and quantification of changes in enzyme activity in live cells. Using our probes, we found the activity of HTRA1 to be highly elevated in an AD-like cell-based model. We also observed the active HTRA2 in live cells by using a mitochondrion-targeted probe. We believe that our probes can serve as a useful tool to study the role of human HTRAs in neurodegenerative diseases.
高温需求 A (HTRA) 丝氨酸蛋白酶家族介导蛋白质质量控制。这些蛋白质在包括阿尔茨海默病 (AD) 和帕金森病 (PD) 在内的几种疾病中处理错误折叠的蛋白质。虽然它们的结构和激活机制已经被研究过，但在生理条件下它们的活性调节的精确细节尚未完全阐明，部分原因是缺乏合适的化学探针。在本研究中，我们开发了针对 HTRA 的新型基于活性的探针 (ABP)，并证明了它们在监测和定量活细胞中酶活性变化方面的实用性。使用我们的探针，我们发现 HTRA1 的活性在类似 AD 的基于细胞的模型中高度升高。我们还通过使用靶向线粒体的探针观察到活细胞中的活性 HTRA2。我们相信我们的探针可以作为研究人类 HTRA 在神经退行性疾病中的作用的有用工具。
