Ma Fei-He, Chen Jia-Liang, Li Qing-Feng, Zuo Hui-Hui, Huang Feng, Su Xun-Cheng
State Key Laboratory of Elemento-organic Chemistry, Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Nankai University, Weijin Road 94, Tianjin 300071 (China), Fax: (+86) 22-23500623.
Chem Asian J. 2014 Jul;9(7):1808-16. doi: 10.1002/asia.201402095. Epub 2014 May 21.
The chemical modification of proteins is a valuable technique in understanding the functions, interactions, and dynamics of proteins. Reactivity and selectivity are key issues in current chemical modification of proteins. The Michael addition-like thiol-ene reaction is a useful tool that can be used to tag proteins with high selectivity for the solvent-exposed thiol groups of proteins. To obtain insight into the bioconjugation of proteins with this method, a kinetic analysis was performed. New vinyl-substituted pyridine derivatives were designed and synthesized. The reactivity of these vinyl tags with L-cysteine was evaluated by UV absorption and high-resolution NMR spectroscopy. The results show that protonation of pyridine plays a key role in the overall reaction rates. The kinetic parameters were assessed in protein modification. The different reactivities of these vinyl tags with solvent-exposed cysteine is valuable information in the selective labeling of proteins with multiple functional groups.
蛋白质的化学修饰是理解蛋白质功能、相互作用和动力学的一项重要技术。反应活性和选择性是当前蛋白质化学修饰中的关键问题。类迈克尔加成硫醇-烯反应是一种有用的工具，可用于对蛋白质溶剂暴露的硫醇基团进行高选择性标记。为深入了解用此方法进行蛋白质生物共轭，进行了动力学分析。设计并合成了新的乙烯基取代吡啶衍生物。通过紫外吸收和高分辨率核磁共振光谱评估了这些乙烯基标签与L-半胱氨酸的反应活性。结果表明，吡啶的质子化在整个反应速率中起关键作用。在蛋白质修饰中评估了动力学参数。这些乙烯基标签与溶剂暴露的半胱氨酸的不同反应活性是用多个官能团选择性标记蛋白质的有价值信息。
