Hoffman Nolan J, Parker Benjamin L, Chaudhuri Rima, Fisher-Wellman Kelsey H, Kleinert Maximilian, Humphrey Sean J, Yang Pengyi, Holliday Mira, Trefely Sophie, Fazakerley Daniel J, Stöckli Jacqueline, Burchfield James G, Jensen Thomas E, Jothi Raja, Kiens Bente, Wojtaszewski Jørgen F P, Richter Erik A, James David E
Charles Perkins Centre, School of Molecular Bioscience, The University of Sydney, Sydney, NSW 2006, Australia.
Garvan Institute of Medical Research, Darlinghurst, NSW 2010, Australia.
Cell Metab. 2015 Nov 3;22(5):922-35. doi: 10.1016/j.cmet.2015.09.001. Epub 2015 Oct 1.
Exercise is essential in regulating energy metabolism and whole-body insulin sensitivity. To explore the exercise signaling network, we undertook a global analysis of protein phosphorylation in human skeletal muscle biopsies from untrained healthy males before and after a single high-intensity exercise bout, revealing 1,004 unique exercise-regulated phosphosites on 562 proteins. These included substrates of known exercise-regulated kinases (AMPK, PKA, CaMK, MAPK, mTOR), yet the majority of kinases and substrate phosphosites have not previously been implicated in exercise signaling. Given the importance of AMPK in exercise-regulated metabolism, we performed a targeted in vitro AMPK screen and employed machine learning to predict exercise-regulated AMPK substrates. We validated eight predicted AMPK substrates, including AKAP1, using targeted phosphoproteomics. Functional characterization revealed an undescribed role for AMPK-dependent phosphorylation of AKAP1 in mitochondrial respiration. These data expose the unexplored complexity of acute exercise signaling and provide insights into the role of AMPK in mitochondrial biochemistry.
运动对于调节能量代谢和全身胰岛素敏感性至关重要。为了探索运动信号网络，我们对未受过训练的健康男性在单次高强度运动前后的人体骨骼肌活检样本中的蛋白质磷酸化进行了全面分析，结果显示在562种蛋白质上有1004个独特的运动调节磷酸化位点。这些位点包括已知的运动调节激酶（AMPK、PKA、CaMK、MAPK、mTOR）的底物，但此前大多数激酶和底物磷酸化位点并未被认为与运动信号有关。鉴于AMPK在运动调节代谢中的重要性，我们进行了有针对性的体外AMPK筛选，并利用机器学习来预测运动调节的AMPK底物。我们使用靶向磷酸蛋白质组学验证了包括AKAP1在内的8种预测的AMPK底物。功能表征揭示了AKAP1的AMPK依赖性磷酸化在线粒体呼吸中具有未被描述的作用。这些数据揭示了急性运动信号尚未被探索的复杂性，并为AMPK在线粒体生物化学中的作用提供了见解。
