新加坡国立大学Shao Q. Yao团队报道了哺乳动物细胞中小分子诱导的翻译后乙酰化催化赖氨酸激酶。相关研究成果发表在2024年8月20日出版的国际知名学术期刊《美国化学会杂志》。

可逆赖氨酸乙酰化是一种重要的翻译后修饰（PTM）。细胞中的这一过程通常由赖氨酸乙酰转移酶和脱乙酰酶酶酶促进行。人激酶中的催化赖氨酸具有高度保守性和可连接性。因此，小分子策略能够以靶向选择性的方式实现激酶上催化赖氨酸的翻译后乙酰化，在激酶生物学中具有巨大的潜力。

该文中，研究人员报告了能够使哺乳动物细胞激酶上的催化赖氨酸进行全局乙酰化的第一种小分子诱导化学策略。通过研究安装在混杂激酶结合支架上的各种赖氨酸乙酰化剂，鉴定出Ac4，并证明其能有效乙酰化活Jurkat/K562细胞中大于100种不同蛋白激酶的催化赖氨酸。为了证明该策略能够靶向蛋白激酶的选择性和可逆化学乙酰化，研究人员在VX-680（AURKA的非共价抑制剂）的基础上进一步开发了六种乙酰化化合物。

其中，Ac13/Ac14虽然显示出优异的体外效力和对AURKA的持续细胞活性，但以靶向选择性的方式显示出其催化赖氨酸（K162）的强烈乙酰化，导致内源性激酶活性的不可逆抑制。在Ac14处理的重组AURKA蛋白上证实了这种化学乙酰化的可逆性，随后用SIRT3（赖氨酸脱乙酰酶）进行脱乙酰化。最后，在SIRT3转染的HCT116细胞中证明了Ac13诱导的内源性AURKA的可逆乙酰化。通过公开第一种能够对激酶上的催化赖氨酸，进行全局和靶向选择性翻译后乙酰化的细胞活性乙酰化化合物，该策略可以为激酶生物学和药物发现提供有用的化学工具。

附：英文原文

Title: Small Molecule-Induced Post-Translational Acetylation of Catalytic Lysine of Kinases in Mammalian Cells

Author: Guanghui Tang, Xuan Wang, Huisi Huang, Manyi Xu, Xingyu Ma, Fengfei Miao, Xiaoyun Lu, Chong-Jing Zhang, Liqian Gao, Zhi-Min Zhang, Shao Q. Yao

Issue&Volume: August 20, 2024

Abstract: Reversible lysine acetylation is an important post-translational modification (PTM). This process in cells is typically carried out enzymatically by lysine acetyltransferases and deacetylases. The catalytic lysine in the human kinome is highly conserved and ligandable. Small-molecule strategies that enable post-translational acetylation of the catalytic lysine on kinases in a target-selective manner therefore provide tremendous potential in kinase biology. Herein, we report the first small molecule-induced chemical strategy capable of global acetylation of the catalytic lysine on kinases from mammalian cells. By surveying various lysine-acetylating agents installed on a promiscuous kinase-binding scaffold, Ac4 was identified and shown to effectively acetylate the catalytic lysine of >100 different protein kinases from live Jurkat/K562 cells. In order to demonstrate that this strategy was capable of target-selective and reversible chemical acetylation of protein kinases, we further developed six acetylating compounds on the basis of VX-680 (a noncovalent inhibitor of AURKA). Among them, Ac13/Ac14, while displaying excellent in vitro potency and sustained cellular activity against AURKA, showed robust acetylation of its catalytic lysine (K162) in a target-selective manner, leading to irreversible inhibition of endogenous kinase activity. The reversibility of this chemical acetylation was confirmed on Ac14-treated recombinant AURKA protein, followed by deacetylation with SIRT3 (a lysine deacetylase). Finally, the reversible Ac13-induced acetylation of endogenous AURKA was demonstrated in SIRT3-transfected HCT116 cells. By disclosing the first cell-active acetylating compounds capable of both global and target-selective post-translational acetylation of the catalytic lysine on kinases, our strategy could provide a useful chemical tool in kinase biology and drug discovery.

DOI: 10.1021/jacs.4c07181

Source: https://pubs.acs.org/doi/abs/10.1021/jacs.4c07181