荷兰内梅亨大学的Wilhelm T. S. Huck课题组近日取得一项新成果。经过不懈努力，他们揭示了可逆的光开关抑制剂产生超灵敏度的非平衡酶反应。 这一研究成果发表在2021年4月12日出版的国际学术期刊《美国化学会杂志》上。

该研究组开发了一种通用的和模块化的策略来使用光可逆地调节酶的活性，并展示了在简单的非平衡酶系统中，当引入可逆的光转换抑制剂(PIs)时，如何产生超灵敏度。利用发色团/弹头策略，合成了蛋白酶α-糜蛋白酶的PIs，发现了对不同光异构体抑制常数(Ki)有很大差异的抑制剂。

使用微流体流动装置，通过连续加入和去除试剂研究非平衡条件下的酶促反应。连续搅拌槽式反应器以不同光脉冲序列照射，即改变紫外光和蓝光照射的脉冲时间或频率，光异构体之间的可逆切换导致酶活性的超灵敏反应和输入信号的频率滤波。

这种通用的和模块化的策略能够对单个酶催化反应的动力学速率进行可逆和可调的控制，并使酶与广泛的网络拓扑结构的可编程连接成为可能。

据介绍，超敏性是生物化学反应网络中普遍存在的一种突发性特性。设计和构建具有超灵敏度的合成反应网络一直具有挑战性，但这将极大地扩展类生命材料的潜在性能。

附：英文原文

Title: Reversible Photoswitchable Inhibitors Generate Ultrasensitivity in Out-of-Equilibrium Enzymatic Reactions

Author: Michael Teders, Aleksandr A. Pogodaev, Glenn Bojanov, Wilhelm T. S. Huck

Issue&Volume: April 12, 2021

Abstract: Ultrasensitivity is a ubiquitous emergent property of biochemical reaction networks. The design and construction of synthetic reaction networks exhibiting ultrasensitivity has been challenging, but would greatly expand the potential properties of life-like materials. Herein, we exploit a general and modular strategy to reversibly regulate the activity of enzymes using light and show how ultrasensitivity arises in simple out-of-equilibrium enzymatic systems upon incorporation of reversible photoswitchable inhibitors (PIs). Utilizing a chromophore/warhead strategy, PIs of the protease α-chymotrypsin were synthesized, which led to the discovery of inhibitors with large differences in inhibition constants (Ki) for the different photoisomers. A microfluidic flow setup was used to study enzymatic reactions under out-of-equilibrium conditions by continuous addition and removal of reagents. Upon irradiation of the continuously stirred tank reactor with different light pulse sequences, i.e., varying the pulse duration or frequency of UV and blue light irradiation, reversible switching between photoisomers resulted in ultrasensitive responses in enzymatic activity as well as frequency filtering of input signals. This general and modular strategy enables reversible and tunable control over the kinetic rates of individual enzyme-catalyzed reactions and makes a programmable linkage of enzymes to a wide range of network topologies feasible.

DOI: 10.1021/jacs.0c12956

Source: https://pubs.acs.org/doi/10.1021/jacs.0c12956