Quantification of exercise‐regulated ubiquitin signaling in human skeletal muscle identifies protein modification cross talk via NEDDylation
Quantification of exercise‐regulated ubiquitin signaling in human skeletal muscle identifies protein modification cross talk via NEDDylation
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DOI：
10.1096/fj.202000075r
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发表时间：
2020-03
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影响因子：
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通讯作者：
B. Parker;B. Kiens;J. Wojtaszewski;E. Richter;David E. James
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作者：
B. Parker;B. Kiens;J. Wojtaszewski;E. Richter;David E. James
维持肌肉功能对全身健康极为重要，而运动对这一过程至关重要。泛素-蛋白酶体系统（UPS）是运动后肌肉适应所必需的，但对高强度运动期间和之后的急性翻译后调节和蛋白质组重塑知之甚少。在这里，我们使用定量蛋白质组学来研究健康男性在单次高强度运动之前，期间和之后的人类骨骼肌活检中的泛素信号动力学。运动导致股外侧肌中蛋白质泛素化的显著耗竭，这与蛋白酶体激活一致。这也与蛋白质丰度的急性翻译后修饰有关。将这些数据与磷酸化蛋白质组学相结合，发现了共调节的近端修饰，表明磷酸化和泛素化之间存在交叉作用。我们还确定了额外的蛋白质修饰串扰，并显示了蛋白质NEDD化的急性激活。体外实验表明，蛋白酶体的cAMP依赖性激活需要NEDD 8（一种参与蛋白质NEDD化的泛素样蛋白）来维持细胞蛋白质泛素化水平。我们的数据揭示了高强度运动期间和之后肌肉中泛素信号传导和蛋白质组重塑的复杂性。我们提出了一个模型，运动和由此产生的cAMP信号通过NEDD化激活蛋白酶体和泛素化，以快速去除潜在的受损蛋白。
The maintenance of muscle function is extremely important for whole body health and exercise is essential to this process. The ubiquitin‐proteasome system (UPS) is required for muscle adaptation following exercise but little is known about acute posttranslational regulation and proteome remodeling during and after high‐intensity exercise. Here, we used quantitative proteomics to study ubiquitin signaling dynamics in human skeletal muscle biopsies from healthy males before, during, and after a single bout of high‐intensity exercise. Exercise resulted in a marked depletion of protein ubiquitylation in the vastus lateralis muscle consistent with proteasome activation. This was also associated with acute posttranslational modification of protein abundance. Integration of these data with phosphoproteomics identified co‐regulated proximal modifications suggesting a cross talk between phosphorylation and ubiquitylation. We also identified additional protein modification cross talk and showed acute activation of protein NEDDylation. In vitro experiments revealed that cAMP‐dependent activation of the proteasome requires NEDD8, an ubiquitin‐like protein involved in protein NEDDylation, to maintain cellular protein ubiquitylation levels. Our data reveal the complexity of ubiquitin signaling and proteome remodeling in muscle during and after high‐intensity exercise. We propose a model whereby exercise and the resulting cAMP signaling activate both the proteasome and ubiquitylation via NEDDylation to rapidly remove potentially damaged proteins.