基于红光驱动的偶氮苯衍生物分子开关光调控蛋溶菌酶构象的研究

This project caters to the need for the integration of technology between chemical analysis and biomedical engineering in our country. We are seeking to find strategies for the photo-control of protein based on the development of photoswitches in biological applications to complementarily figure out how to make the control effectively on the wavelengths with tissue penetration. First, egg white lysozyme was selected as the target protein due to its natural properties and azobenzene-containing photoswitches driven by red light is required to design in this experiment, thereby establishing an empirical model of azobenzene cross-linker position on the structure of lysozyme. Subsequently, attempts will be conducted to reveal the binding mechanism of the azobenzene derivative with functionalized groups and lysozyme via computational emulation, modern measurement technology, and morphological analysis. Furthermore, this work will focus on photo-controlling conformations of lysozyme in a reversible (on/off) manner with red light-controlled azobenzene derivative switches and optimizing the chemical process development of photoresponsive functional devices. The results will provide theoretical basis and technical reserves for real-time monitoring and controlling of the biological reconstruction, especially for the therapy of protein conformational disease.

本项目面向我国化学分析技术与生物医学工程交叉融合的发展需求，在前期开发光分子开关的工作基础之上，进一步研究光调控蛋白质策略，辅助解决该策略难以在与生物组织高度相容的波长范围得到有效控制的科学问题。实验选取自然存在于鸡蛋清中的蛋溶菌酶作为光调控致病蛋白构象变化的模型蛋白，从设计合成可用红光驱动的偶氮苯衍生物分子开关为切入点，建立偶氮分子化学修饰蛋溶菌酶结构的交联模型；借助计算机模拟仿真、现代测试技术和形态学分析手段，揭示含特定官能团的偶氮苯衍生物与蛋溶菌酶的作用机理；开发基于偶氮苯衍生物分子开关红光区有效调控蛋溶菌酶构象的可逆光化学操作策略，优化和改进光响应元件的开发工艺，为直接检测和实时调控生物体内有蛋白质参与的生物学过程及蛋白质构象病的治疗提供理论基础和技术储备。

本项目设计和优化了具有合适端到端距离和顺-反光致异构化特性的马来酰亚胺偶氮苯衍生物，探讨了基于马来酰亚胺基团的偶氮苯衍生物在不同光源条件下的结构特性，合成了一种可用红光驱动的偶氮苯衍生物分子开关。该光开关分子在625nm 红光充分照射下可达到99%以上的构型转化。实验中，阐明了含马来酰亚胺基团的偶氮苯衍生物与蛋溶菌酶的作用机制，建立了偶氮苯衍生物-蛋溶菌酶体外结合模型，确定了含特殊基团红光驱动的偶氮苯衍生物分子开关交联蛋溶菌酶结构的化学方式。热弛豫数据表明偶氮分子开关交联蛋白质后的顺反异构转化时间会延长且交联反应会生成含琥珀亚酰胺的水解副产物，但不影响其对多肽及蛋白质的构象调控。研究中，揭示了蛋溶菌酶构象受偶氮苯衍生物化学修饰的光学特性和构象改变，并基于对比模型评价了红光驱动的偶氮苯衍生物分子开关光调控蛋溶菌酶的可行性，证明了调控的有效性。该项目成果辅助解决了光调控策略难以在与生物组织高度相容的波长范围得到有效控制的科学问题，为光调控天然蛋白质的科学研究开拓思路。