微管是真核细胞中普遍存在的细胞骨架成分之一，具有重要的生理生化功能，同时也是抗癌药物的重要作用靶点之一。控制细胞中的微管调节机制即为“微管蛋白密码（Tubulin code）”，主要包括同型（Isotype）微管蛋白种类的多样性和微管蛋白翻译后修饰的复杂性。目前对各种同型微管蛋白的功能研究较少。我们以β微管蛋白TUBB3为研究对象，以长春碱为β微管蛋白的配体，采用Bump-Hole化学遗传学技术，从同型微管蛋白种类的多样性方面进行“微管蛋白密码”的解码研究。首先，通过分子对接和体外的功能验证筛选出一对生物正交的突变TUBB3和长春碱类似物，在正常细胞、肿瘤细胞和神经元细胞中建立生物正交体系。然后用这些配体处理细胞，通过比较生物正交体系和非正交体系以及未用配体处理的体系的转录组和蛋白组差异，探究TUBB3参与的生物学过程，系统的揭示TUBB3的功能，为解析“微管蛋白密码”提供新思路和新见解。

As the major component of cytoskeleton, microtubule possesses essential physiological and biochemical functions. It is also an important target of anti-tumor drugs. A regulatory mechanism to control the specialization of the microtubule cytoskeleton is the “tubulin code”, which is generated by expression of different α- and β-tubulin isotypes, and by post-translational modifications of tubulin. So far, little is known regarding the impact of specific tubulin isotypes on MT properties and functions. Bump-Hole strategy may address this problem. Firstly, modified vinblastine and mutant human βIII-tubulin (TUBB3) were designed according to the co-crystal structure of microtubule and vinblastine, and one set of bioorthogonal systems were established based on Rosetta ligand docking, in vitro function analysis. Then, wild type and mutant cells were treated with unmodified or modified vinblastine or DMSO. Through analyzing the differences of transcriptomes and proteomes of these treatments, we can further understand the functions of TUBB3, which will provide new ideas and insights in revealing the functions of different tubulin isotypes and deciphering the “tubulin code”.