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使用SpyTag和阿尔科西尔酸进行pH依赖的SpyCatcher/Tag系统激活

Sonji Che¹, Hiroyuki Konno¹, Koki Makabe¹

¹Graduate School of Science and Engineering, Yamagata University, 4-3-16 Jyonan, Yonezawa, Yamagata 992-8510, Japan.

Bioconjugate chemistry

|April 26, 2024

概括

这项研究修改了SpyTag用于pH激活蛋白结合. 一种新的Pro修改可以防止不必要的异酸盐的形成,从而实现受控的蛋白质工程应用.

科学领域:

生物化学生物化学
蛋白质工程是指蛋白质的工程.
合成生物学合成生物学

背景情况:

通过SpyCatcher/SpyTag系统,可以在没有外部能量来源的情况下进行共价蛋白质结合.
该系统的条件激活可以提高其在蛋白质工程中的效用.
研究Alkoxyaspartic酸用于pH触发的SpyTag激活.

研究的目的:

为了评估使用alkoxyaspartic酸SpyTag的pH激活.
为了解决高pH激活过程中异酸盐形成的问题.
开发一种经过修改的SpyTag,具有受控激活.

主要方法:

在SpyTag.的异酸形成残留物中加入alkoxyaspartic酸.
根据pH值进行的激活和水解研究.
修改C端酸位置的变化为proline.

主要成果:

一个具有乙氧基的体通过水解显示了pH的激活.
水解导致了不必要的异酸盐 (isoAsp) 的形成.
将C终端的酸替换为proline,可以在高pH下防止isoAsp的形成.

结论:

氧酸的修饰使pH触发的SpyTag激活成为可能.
林替代是一种可行的策略,可以防止isoAsp的形成.
这项工作促进了对SpyTag的合成构建,用于受控的应用.

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