Yu Jianjun, Liu Huijie, Gao Rui, Wang Tao V, Li Chenggang, Liu Yuxiang, Yang Lu, Xu Ying, Cui Yunfeng, Jia Chenxi, Huang Juan, Chen Peng R, Rao Yi
Laboratory of Neurochemical Biology, Peking-Tsinghua Center for Life Sciences, Peking-Tsinghua-NIBS (PTN) Graduate Program, School of Life Sciences, Peking University, Beijing, China; Chinese Institute for Brain Research (CIBR), Beijing, China; Department of Chemical Biology, College of Chemistry and Chemical Engineering; School of Pharmaceutical Sciences, PKU-IDG/McGovern Institute for Brain Research, Peking University, Beijing, China; Chinese Institutes for Medical Research (CIMR), Beijing, China; Capital Medical University, Beijing, China.
Laboratory of Neurochemical Biology, Peking-Tsinghua Center for Life Sciences, Peking-Tsinghua-NIBS (PTN) Graduate Program, School of Life Sciences, Peking University, Beijing, China; Chinese Institute for Brain Research (CIBR), Beijing, China; Department of Chemical Biology, College of Chemistry and Chemical Engineering; School of Pharmaceutical Sciences, PKU-IDG/McGovern Institute for Brain Research, Peking University, Beijing, China.
Cell Chem Biol. 2025 Jan 16;32(1):157-173.e7. doi: 10.1016/j.chembiol.2024.12.003. Epub 2024 Dec 30.
Research into mechanisms underlying sleep traditionally relies on electrophysiology and genetics. Because sleep can only be measured on whole animals by behavioral observations and physical means, no sleep research was initiated by biochemical and chemical biological approaches. We used phosphorylation sites of kinases important for sleep as targets for biochemical and chemical biological approaches. Sleep was increased in mice carrying a threonine-to-alanine substitution at residue T469 of salt-inducible kinase 3 (SIK3). Our biochemical purification and photo-crosslinking revealed calcineurin (CaN) dephosphorylation, both in vitro and in vivo, of SIK3 at T469 and S551, but not T221. Knocking down CaN regulatory subunit reduced daily sleep by more than 5 h, exceeding all known mouse mutants. Our work uncovered a critical physiological role for CaN in sleep and pioneered biochemical purification and chemical biology as effective approaches to study sleep.
传统上，对睡眠潜在机制的研究依赖于电生理学和遗传学。由于睡眠只能通过行为观察和物理手段在完整动物身上进行测量，因此尚未有通过生化和化学生物学方法开展的睡眠研究。我们将对睡眠至关重要的激酶磷酸化位点作为生化和化学生物学方法的靶点。在盐诱导激酶3（SIK3）的第469位残基处携带苏氨酸到丙氨酸替换的小鼠中，睡眠时间增加。我们的生化纯化和光交联实验表明，在体外和体内，钙调神经磷酸酶（CaN）均可使SIK3的第469位和第551位残基去磷酸化，但不能使第221位残基去磷酸化。敲低CaN调节亚基可使每日睡眠时间减少超过5小时，超过所有已知的小鼠突变体。我们的工作揭示了CaN在睡眠中的关键生理作用，并开创了生化纯化和化学生物学作为研究睡眠的有效方法。
