ACS Chem Biol. 2019 Oct 18;14(10):2276-2285. doi: 10.1021/acschembio.9b00614. Epub 2019 Sep 25.
Imaging of an active protease with an exquisite specificity in the presence of highly homologous proteins within a living cell is a very challenging task. Herein, we disclose a new method called "Activity-based Reporter Gene Technology" (AbRGT). This method provides an opportunity to study the function of "active protease" with an unprecedented specificity. As a proof-of-concept, we have applied this method to study the function of individual caspase protease in both intrinsic and extrinsic apoptosis signaling pathways. The versatility of this method is demonstrated by studying the function of both the initiator and effector caspases, independently. The modular fashion of this technology provides the opportunity to noninvasively image the function of cathepsin-B in a caspase-dependent cell death pathway. As a potential application, this method is used as a tool to screen compounds that are potent inhibitors of caspases and cathepsin-B proteases. The fact that this method can be readily applied to any protease of interest opens up huge opportunities for this technology in the area of target validation, high-throughput screening, imaging, diagnostics, and therapeutic intervention.
在活细胞内，针对高度同源的蛋白质，对活性蛋白酶进行成像，这是一项极具挑战性的任务。在此，我们揭示了一种新方法，称为“基于活性的报告基因技术”（AbRGT）。该方法为研究“活性蛋白酶”的功能提供了前所未有的特异性。作为概念验证，我们已经将该方法应用于研究内在和外在细胞凋亡信号通路中单个半胱天冬酶蛋白酶的功能。该方法的多功能性通过独立研究起始半胱天冬酶和效应半胱天冬酶的功能得到证明。该技术的模块化设计为非侵入性地研究半胱天冬酶依赖性细胞死亡途径中组织蛋白酶-B 的功能提供了机会。作为一种潜在的应用，该方法可用作筛选有效抑制半胱天冬酶和组织蛋白酶-B 蛋白酶的化合物的工具。事实上，该方法可以轻松应用于任何感兴趣的蛋白酶，这为该技术在靶标验证、高通量筛选、成像、诊断和治疗干预等领域开辟了巨大的机会。
