selectivity. A phage-display screen of a mutant sortase library that was randomized in the substrate recognition loop was used to isolate this mutant. The altered substrate selectivity represents a gain-of-function that was exploited for the traceless semisynthesis of histone H3. Our report is a decisive step toward a platform of engineered sortases with distinct ligation properties that will conceivably
细菌转肽酶分选酶 A 以
化学选择性方式连接两个
多肽已成为蛋白质工程方法的通用工具。当分选酶介导的连接用于蛋白质半合成时,由于 L
PxTG 分选基序的严格要求而导致的多达四个突变被引入目标蛋白质。在这里,我们报告了具有广泛底物选择性的突变分选酶 A 的定向进化。使用在底物识别环中随机化的突变分选酶文库的噬菌体展示屏幕来分离该突变体。改变的底物选择性代表了一种功能增益,可用于组蛋白 H3 的无痕半合成。