Proteomic analysis revealed different responses to hypergravity of soleus and extensor digitorum longus muscles in mice
Proteomic analysis revealed different responses to hypergravity of soleus and extensor digitorum longus muscles in mice
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DOI：
10.1016/j.jprot.2020.103686
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发表时间：
2020-04-15
影响因子：
3.3
通讯作者：
Hirano, Hisashi
中科院分区：
文献类型：
--
作者：
Ohira, Takashi;Ino, Yoko;Hirano, Hisashi
研究蛋白质丰度谱对于了解慢骨骼肌和快骨骼肌特征的差异非常重要。比较了1g和3g染毒28 d小鼠比目鱼肌和趾长伸肌的形态学变化。这些图谱的生物学意义表明，超重力暴露激活了大量参与溶胶中蛋白质合成的途径。相反，参与氧化磷酸化的信号通路的失活在EDL中是明显的。这些结果表明，在溶胶和EDL分子途径的反应性不同。此外，亚精胺合酶和亚精胺，细胞生长的一个重要的多胺的水平，增加在两个肌肉超重力暴露后，而精胺氧化酶（SMOX）的水平增加在EDL单独。SMOX水平与参与肌肉萎缩的精胺含量呈负相关，并且在EDL中高于Sol，即使在1 g组中也是如此。这些结果表明，SMOX对Sol和EDL中精脒和精胺含量的调节作用不同。然而，与预期相反，SMOX水平的差异并没有对这些肌肉的生长产生显着影响以下hypergravityexposure.Significance：骨骼肌特异性蛋白质丰度分布导致慢骨骼肌和快骨骼肌的特性差异。我们通过LC-MS/MS分析和无标记定量研究了小鼠慢收缩Sol和快收缩EDL肌肉在1 g和3 g暴露28天后的特征差异。骨骼肌蛋白的两步溶解增加了通过LC-MS/MS分析鉴定的蛋白的覆盖率。此外，该方法比通过SDS-PAGE和离线HPLC的蛋白质或肽分级分离更容易降低样品的复杂性，同时保持样品的高可操作性，并且具有重现性。大量的超重力反应蛋白，以及显着增加湿重观察溶胶比EDL肌肉。1g和3g组蛋白质丰度谱差异的生物学意义表明，Sol和EDL肌肉中每种分子途径对超重力暴露的反应性差异很大。此外，我们发现哺乳动物细胞生长和生存的重要因素多胺的生物合成和相互转化途径对超重力暴露有反应;即使在1g组中，Sol和EDL肌肉中的亚精胺和精胺含量也受到不同机制的调节。然而，我们的研究结果表明，调节多胺含量的机制的差异是不可能有显着的影响，溶胶和EDL肌肉生长的差异，超重力暴露。
Investigating protein abundance profiles is important to understand the differences in the slow and fast skeletal muscle characteristics. The profiles in soleus (Sol) and extensor digitorum longus (EDL) muscles in mice exposed to 1 g or 3 g for 28 d were compared. The biological implications of the profiles revealed that hypergravity exposure activated a larger number of pathways involved in protein synthesis in Sol. In contrast, the inactivation of signalling pathways involved in oxidative phosphorylation were conspicuous in EDL. These results suggested that the reactivity of molecular pathways in Sol and EDL differed. Additionally, the levels of spermidine synthase and spermidine, an important polyamine for cell growth, increased in both muscles following hypergravity exposure, whereas the level of spermine oxidase (SMOX) increased in EDL alone. The SMOX level was negatively correlated with spermine content, which is involved in muscle atrophy, and was higher in EDL than Sol, even in the 1 g group. These results indicated that the contribution of SMOX to the regulation of spermidine and spermine contents in Sol and EDL differed. However, contrary to expectations, the difference in the SMOX level did not have a significant impact on the growth of these muscles following hypergravity exposure.Significance: The skeletal muscle-specific protein abundance profiles result in differences in the characteristics of slow and fast skeletal muscles. We investigated differences in the profiles in mouse slow-twitch Sol and fasttwitch EDL muscles following 28-d of 1 g and 3 g exposure by LC-MS/MS analysis and label-free quantitation. A two-step solubilisation of the skeletal muscle proteins increased the coverage of proteins identified by LC-MS/MS analysis. Additionally, this method reduced the complexity of samples more easily than protein or peptide fractionation by SDS-PAGE and offline HPLC while maintaining the high operability of samples and was reproducible. A larger number of hypergravity-responsive proteins as well as a prominent increase in the wet weights was observed in Sol than EDL muscles. The biological implications of the difference in the protein abundance profiles in 1 g and 3 g groups revealed that the reactivity of each molecular pathway in Sol and EDL muscles to hypergravity exposure differed significantly. In addition, we found that the biosynthetic and interconversion pathway of polyamines, essential factors for cell growth and survival in mammals, was responsive to hypergravity exposure; spermidine and spermine contents in Sol and EDL muscles were regulated by different mechanisms even in the 1 g group. However, our results indicated that the difference in the mechanism regulating polyamine contents is unlikely to have a significant effect on the differences in Sol and EDL muscle growth following hypergravity exposure.