Institut de Recerca Biomèdica, 08028 Barcelona, Spain.
Chembiochem. 2009 Sep 21;10(14):2361-6. doi: 10.1002/cbic.200900244.
Postproline proteases constitute a subset of serine proteases involved in the regulation of many signaling events and are emerging as promising therapeutic targets for prevalent diseases, such as diabetes and cancer. Therefore, monitoring their activity in different tissues and diverse physiological states would certainly facilitate the elucidation of their physiological role and the establishment of new therapeutic targets. Here, we have synthesized a dipeptidyl phosphonate activity-based probe that has proved to be highly selective for a specific postproline protease, prolyl oligopeptidase (POP). Its high sensitivity allows the detection of the endogenous activity of POP both by in-gel analysis and mass spectrometry. The evidence provided by mass spectrometry for the high selectivity of the synthesized probe opens the possibility of using dipeptidyl phosphonates not only for activity-based profiling (ABP), but also for other ABP applications like substrate-based protease identification.
脯氨酰肽酶构成了丝氨酸蛋白酶的一个亚类，参与许多信号事件的调节，并且作为治疗常见疾病（如糖尿病和癌症）的有前途的靶点而出现。因此，监测它们在不同组织和不同生理状态下的活性肯定会有助于阐明它们的生理作用，并建立新的治疗靶点。在这里，我们合成了一种二肽基膦酸活性探针，该探针已被证明对特定的脯氨酰肽酶，脯氨酰寡肽酶（POP）具有高度选择性。其高灵敏度允许通过凝胶分析和质谱法检测内源性 POP 活性。质谱法提供的证据表明，所合成探针具有高选择性，这为使用二肽基膦酸盐不仅进行基于活性的蛋白质组学分析（ABP），而且为其他基于底物的蛋白酶鉴定等 ABP 应用开辟了可能性。
