基于邻近的分选酶介导连接（PBSL）对重组蛋白进行位点特异性C末端标记
Site-Specific C-Terminal Labeling of Recombinant Proteins with Proximity-Based Sortase-Mediated Ligation (PBSL).
作者信息
Wang Hejia Henry, Tsourkas Andrew
机构信息
Biochemistry and Molecular Biophysics Graduate Group, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA.
出版信息
Methods Mol Biol. 2019;2012:15-28. doi: 10.1007/978-1-4939-9546-2_2.
S. aureus sortase A (SrtA), a calcium-dependent transpeptidase, is frequently employed to site-specifically label the C-terminus of recombinant proteins bearing an LPXTG SrtA recognition motif. Unfortunately, SrtA suffers from low turnover rates, resulting in poor ligation efficiencies even with optimized reaction conditions. In this chapter, we describe proximity-based sortase-mediated ligation (PBSL), which uses the SpyTag-SpyCatcher peptide-protein pair to link SrtA to target proteins and dramatically improves reaction rate and ligation efficiency.
金黄色葡萄球菌分选酶A（SrtA）是一种钙依赖性转肽酶，常用于对带有LPXTG SrtA识别基序的重组蛋白的C末端进行位点特异性标记。不幸的是，SrtA的周转速率较低，即使在优化的反应条件下，连接效率也很差。在本章中，我们描述了基于邻近性的分选酶介导的连接（PBSL），它使用SpyTag-SpyCatcher肽-蛋白对将SrtA与靶蛋白连接起来，从而显著提高反应速率和连接效率。
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