蛋氨酸亚砜还原酶荧光探针的设计及应用

A novel fluorescent probe for methionine sulfoxide reductases

蛋氨酸亚砜还原酶(Msr)是一类很好的抗氧化酶和蛋白修复酶,在生物体氧化还原体系中扮演着重要的角色.本文依据其反应特异性及分子探针发光机制,设计合成了可用于检测蛋氨酸亚砜还原酶活性的探针Msr-TFMCM,并详细研究了其与Msrs的相互作用及反应机理.在此基础上,将Msr-TFMCM进一步用于活细胞内Msrs活性的可视化检测.在培养的细胞中预先加入Msrs的底物蛋氨酸亚砜可以显著抑制荧光信号,说明了探针可以实现对Msrs的专一性检测.

Methionine sulfoxide reductase （Msr） enzymes have multiple functions such as antioxidant defense and protein repair, and play an important role in regulating cellular redox homeostasis. However, the investigation of the enzymes’ function under physiological/pathological conditions is challenging as there are only limited probes available.In this work, we employed the trifluoromethyl coumarin as a fluorophore and the intramolecular charge transfer sensing mechanism to design a turn-on fluorescent probe, Msr-TFMCM. The methylsulfoxide group in Msr-TFMCM is specifically reduced by Msrs, giving a 50-fold increase of the emission intensity. The practical applications of the probe were demonstrated by imaging the enzymes’ activity in live cells. Our results indicated that Msr-TFMCM could be used for specific monitoring Msrs activity.