Department of Chemistry, University of Leicester, Leicester, LE1 7RH, UK.
MIP Diagnostics Ltd, Colworth Science Park, Sharnbrook, MK44 1LQ, UK.
J Mater Chem B. 2022 Sep 15;10(35):6732-6741. doi: 10.1039/d2tb00278g.
Modulation of enzyme activity allows for control over many biological pathways and while strategies for the pharmaceutical design of inhibitors are well established; methods for promoting activation, that is an increase in enzymatic activity, are not. Here we demonstrate an innovative epitope mapping technique using molecular imprinting to identify four surface epitopes of acetylcholinesterase (AChE). These identified epitopes were then used as targets for the synthesis of molecularly imprinted nanoparticles (nanoMIPs). The enzymatic activity of AChE was increased upon exposure to these nanoMIPs, with one particular identified epitope nanoMIP leading to an increase in activity of 47× compared to enzyme only. The impact of nanoMIPs on the inhibited enzyme is also explored, with AChE activity recovering from 11% (following exposure to an organophosphate) to 73% (following the addition of nanoMIPs). By stabilizing the conformation of the protein rather than targeting the active site, the allosteric nature of MIP-induced reactivation suggests a new way to promote enzyme activity, even under the presence of an inhibitor. This method of enzyme activation shows promise to treat enzyme deficiency diseases or in medical emergencies where an external agent affects protein function.
酶活性的调节可以控制许多生物途径，虽然抑制剂的药物设计策略已经成熟，但促进激活（即增加酶活性）的方法却没有。在这里，我们展示了一种使用分子印迹技术进行表位作图的创新技术，以鉴定乙酰胆碱酯酶（AChE）的四个表面表位。然后，将这些鉴定的表位用作合成分子印迹纳米颗粒（nanoMIP）的靶标。暴露于这些 nanoMIP 后，AChE 的酶活性增加，其中一个特定鉴定的表位 nanoMIP 导致活性增加了 47 倍，而与仅酶相比。还探索了 nanoMIP 对受抑制酶的影响，AChE 活性从 11%（暴露于有机磷后）恢复到 73%（添加 nanoMIP 后）。通过稳定蛋白质的构象而不是靶向活性位点，MIP 诱导的重新激活的变构性质表明了一种促进酶活性的新方法，即使在存在抑制剂的情况下也是如此。这种酶激活方法有望治疗酶缺乏症或在外部试剂影响蛋白质功能的医疗紧急情况下使用。
