Fan Zhuming, Liu Zhiyang, Zhang Nan, Wei Wenyu, Cheng Ke, Sun Hongyan, Hao Quan
Institute of High Energy Physics, CAS, Beijing 100000, China.
Spallation Neutron Source Science Center, CAS, Dongguan, Guangdong 523000, China.
iScience. 2023 Aug 28;26(10):107757. doi: 10.1016/j.isci.2023.107757. eCollection 2023 Oct 20.
Lysine lactylation (Kla) is a novel histone post-translational modification discovered in late 2019. Later, HDAC1-3, were identified as the robust Kla erasers. While the Sirtuin family proteins showed weak eraser activities toward Kla, as reported. However, the catalytic mechanisms and physiological functions of HDACs and Sirtuins are not identical. In this study, we observed that SIRT3 exhibits a higher eraser activity against the H4K16la site than the other human Sirtuins. Crystal structures revealed the detailed binding mechanisms between lactyl-lysine peptides and SIRT3. Furthermore, a chemical probe, p-H4K16laAlk, was developed to capture potential Kla erasers from cell lysates. SIRT3 was captured by this probe and detected via proteomic analysis. And another chemical probe, p-H4K16la-NBD, was developed to detect the eraser-Kla delactylation processes directly via fluorescence indication. Our findings and chemical probes provide new directions for further investigating Kla and its roles in gene transcription regulation.
赖氨酸乳酰化（Kla）是2019年末发现的一种新型组蛋白翻译后修饰。后来，HDAC1 - 3被鉴定为强效的Kla去修饰酶。虽然据报道，沉默调节蛋白家族蛋白对Kla表现出较弱的去修饰酶活性。然而，HDACs和沉默调节蛋白的催化机制和生理功能并不相同。在本研究中，我们观察到SIRT3对H4K16la位点的去修饰酶活性高于其他人类沉默调节蛋白。晶体结构揭示了乳酰 - 赖氨酸肽与SIRT3之间的详细结合机制。此外，还开发了一种化学探针p - H4K16laAlk，用于从细胞裂解物中捕获潜在的Kla去修饰酶。该探针捕获了SIRT3，并通过蛋白质组学分析进行了检测。另外还开发了一种化学探针p - H4K16la - NBD，用于通过荧光指示直接检测去修饰酶 - Kla去乳酰化过程。我们的研究结果和化学探针为进一步研究Kla及其在基因转录调控中的作用提供了新的方向。
