Academy for Advanced Interdisciplinary Studies, Peking University, Beijing, China; Peking-Tsinghua Center for Life Sciences, Beijing, China.
Beijing National Laboratory for Molecular Sciences, Synthetic and Functional Biomolecules Center, Department of Chemical Biology, College of Chemistry and Molecular Engineering, Peking University , Beijing, China.
ACS Cent Sci. 2016 May 25;2(5):325-31. doi: 10.1021/acscentsci.6b00024. Epub 2016 Apr 27.
Selective manipulation of protein kinases under living conditions is highly desirable yet extremely challenging, particularly in a gain-of-function fashion. Here we employ our recently developed bioorthogonal cleavage reaction as a general strategy for intracellular activation of individual kinases. Site-specific incorporation of trans-cyclooctene-caged lysine in place of the conserved catalytic lysine, in conjunction with the cleavage partner dimethyl-tetrazine, allowed efficient lysine decaging with the kinase activity chemically rescued in living systems.
在活细胞条件下对蛋白质激酶进行选择性操作是非常理想的，但极具挑战性，尤其是以获得功能的方式。在这里，我们采用最近开发的生物正交裂解反应作为一种通用策略，用于在细胞内激活单个激酶。通过将反式环辛烯封闭的赖氨酸定点整合到保守的催化赖氨酸位置，与裂解伴侣二甲四嗪结合，可在活细胞系统中有效去封闭赖氨酸，并通过化学方法恢复激酶活性。
