Yang Zhongguang, Fok Hong Kiu Francis, Luo Jiren, Yang Yang, Wang Ri, Huang Xinyu, Sun Fei
Department of Chemical and Biological Engineering, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China.
Division of Life Science, The Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong SAR, China.
Sci Adv. 2022 Apr;8(13):eabm5482. doi: 10.1126/sciadv.abm5482. Epub 2022 Apr 1.
Although the tools based on split proteins have found broad applications, ranging from controlled biological signaling to advanced molecular architectures, many of them suffer from drawbacks such as background reassembly, low thermodynamic stability, and static structural features. Here, we present a chemically inducible protein assembly method enabled by the dissection of the carboxyl-terminal domain of a B-dependent photoreceptor, CarH. The resulting segments reassemble efficiently upon addition of cobalamin (AdoB, MeB, or CNB). Photolysis of the cofactors such as AdoB and MeB further leads to stable protein adducts harboring a bis-His-ligated B. Split CarH enables the creation of a series of protein hydrogels, of which the mechanics can be either photostrengthened or photoweakened, depending on the type of B. These materials are also well suited for three dimensional cell culturing. Together, this new protein chemistry, featuring negligible background autoassembly, stable conjugation, and phototunability, has opened up opportunities for designing smart materials.
尽管基于分裂蛋白的工具已得到广泛应用，涵盖从可控生物信号传导到先进分子结构等领域，但其中许多工具存在诸如背景重新组装、热力学稳定性低和静态结构特征等缺点。在此，我们提出一种化学诱导蛋白组装方法，该方法通过剖析一种依赖钴胺素的光感受器CarH的羧基末端结构域来实现。加入钴胺素（腺苷钴胺素、甲基钴胺素或氰钴胺素）后，产生的片段能高效重新组装。腺苷钴胺素和甲基钴胺素等辅因子的光解进一步导致含有双组氨酸连接的钴胺素的稳定蛋白加合物。分裂的CarH能够创建一系列蛋白质水凝胶，其力学性能可根据钴胺素的类型进行光增强或光减弱。这些材料也非常适合三维细胞培养。总之，这种新的蛋白质化学具有可忽略不计的背景自动组装、稳定的共轭作用和光可调性等特点，为设计智能材料开辟了机会。
