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使用 CoLDR 化学方法对内源蛋白进行特定地点的标记

Rambabu N Reddi¹, Adi Rogel¹, Efrat Resnick¹

¹Department of Chemical and Structural Biology, The Weizmann Institute of Science, Rehovot, 7610001, Israel.

Journal of the American Chemical Society

|November 24, 2021

概括

一种新的共价配体定向释放 (CoLDR) 方法可以在细胞中进行特定位点的蛋白质标记,而不会抑制蛋白质活性. 这种化学生物学工具可以安装多种功能标签,并保持标蛋白的功能.

科学领域:

化学生物学
分子生物学
生物化学

背景情况:

化学修饰会影响蛋白质功能,但细胞中特定的非遗传标记是有限的.
传统的蛋白质标记方法可以通过占用结合点来抑制蛋白质活性.

研究的目的:

开发一种新的特定区域的蛋白质标记策略,以保持蛋白质的活性.
展示这种新标签方法在细胞环境中的多功能性和应用.

主要方法:

开发了对特定位点蛋白质进行标记的共价配体定向释放 (CoLDR) 策略.
应用 CoLDR 用各种标签标记蛋白质,包括 BTK,K-Ras^G12C和 SARS-CoV-2 PL^pro.
使用 CoLDR 来确定蛋白质半衰期,量化降解,监测配体结合,并开发 PROTAC.

主要成果:

在细胞中用基和基标签对BTK进行选择性标记.
证明 CoLDR 标签可以保持 BTK 活性,与传统方法不同.
通过使用PROTACs成功量化BTK降解,并监测到BTK活性部位的配体结合.

结论:

CoLDR是一种强大的化学生物学工具,用于对特定区域的蛋白质进行标记,从而维持蛋白质的活性.
这种方法为研究原生环境中的蛋白质动力学和相互作用提供了最小的干扰.
CoLDR扩大了化学生物学研究的工具箱,使得新的应用程序,如高效的PROTAC开发成为可能.

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