Wang Xiao-Wei, Zhang Wen-Bin
Beijing National Laboratory for Molecular Sciences, Key Laboratory of Polymer Chemistry and Physics of Ministry of Education, Center for Soft Matter Science and Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing, People's Republic of China.
Methods Mol Biol. 2019;2033:287-300. doi: 10.1007/978-1-4939-9654-4_19.
The emergence of "molecular superglue," such as SpyTag-SpyCatcher chemistry, has tremendously expanded our capability in manipulating protein shape and architecture via conjugation. Telechelic proteins bearing the SpyTag and SpyCatcher reactive sequences can be expressed and purified for bioconjugation in vitro, giving protein conjugates, branched proteins, and circular proteins. By encoding both reactive sequences in the same construct for expression in vivo, the nascent protein undergoes programmed posttranslational modification guided by protein folding and reaction, leading to diverse nonlinear topologies in situ. In this chapter, we present the SpyTag-SpyCatcher chemistry as a versatile platform for protein bioconjugation and topology engineering.
“分子超级胶水”的出现，如SpyTag-SpyCatcher化学，极大地扩展了我们通过共轭操纵蛋白质形状和结构的能力。带有SpyTag和SpyCatcher反应序列的遥爪蛋白可以表达并纯化用于体外生物共轭，从而得到蛋白质共轭物、分支蛋白和环状蛋白。通过在同一构建体中编码这两个反应序列以在体内表达，新生蛋白质会在蛋白质折叠和反应的引导下进行程序性翻译后修饰，从而原位产生多种非线性拓扑结构。在本章中，我们将介绍SpyTag-SpyCatcher化学作为蛋白质生物共轭和拓扑工程的通用平台。
