Myofibrillar Z-discs Are a Protein Phosphorylation Hot Spot with Protein Kinase C (PKCα) Modulating Protein Dynamics*
Myofibrillar Z-discs Are a Protein Phosphorylation Hot Spot with Protein Kinase C (PKCα) Modulating Protein Dynamics*
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肌原纤维 Z 盘是蛋白磷酸化热点，蛋白激酶 C (PKCα) 调节蛋白动力学*
DOI：
10.1074/mcp.m116.065425
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发表时间：
2017
影响因子：
7
通讯作者：
Warscheid B
中科院分区：
文献类型：
--
作者：
Reimann L;Wiese H;Leber Y;Schwäble AN;Rohland A;Knapp B;Peikert CD;Drepper F;van der Ven PFM;Radziwill G;Fürst DO;Warscheid B
Z盘是一种富含蛋白质的结构，对于肌原纤维的发育和完整性至关重要，肌原纤维是横纹肌细胞的收缩细胞器。我们在这里使用小鼠C2C12成肌细胞，其被分化成肌管，随后通过电脉冲刺激（EPS）产生包含成熟Z盘的收缩肌管。使用定量蛋白质组学方法，我们发现成肌细胞与分化的肌管中387种蛋白质的相对丰度发生了显着变化，反映了这些细胞在肌发生过程中的剧烈表型转换。有趣的是，诱导Z盘组装和成熟的分化肌管的EPS导致参与ATP合成的蛋白质水平增加，可能是为了满足收缩肌管的更高能量需求。由于最近提出了Z盘在信号整合和转导中的重要作用，因此其精确的磷酸化景观进一步值得深入分析。因此，我们建立了全球磷酸化蛋白质组学的EPS处理的收缩肌管，一个全面的网站解决的蛋白磷酸化地图的Z盘，并发现它是一个磷酸化热点骨骼肌细胞，强调其功能的信号和疾病相关的过程。在一个说明性的方式，我们分析了肌动蛋白结合的多接头蛋白细丝蛋白C（FLNc），这是必不可少的Z盘组装和维护，并发现PKCα磷酸化在不同的丝氨酸残基在其铰链2区防止其切割在邻近的酪氨酸残基的钙蛋白酶1。光漂白实验后的荧光恢复表明，这种磷酸化调节FLNc动力学。此外，缺乏裂解的Ig样结构域24的FLNc表现出非常快的动力学和极高的迁移率。我们的数据集提供了研究社区资源，用于进一步鉴定肌原纤维蛋白相互作用、动力学和流动性中激酶介导的变化，这将大大促进我们对Z盘动力学和信号传导的理解。
The Z-disc is a protein-rich structure critically important for the development and integrity of myofibrils, which are the contractile organelles of cross-striated muscle cells. We here used mouse C2C12 myoblast, which were differentiated into myotubes, followed by electrical pulse stimulation (EPS) to generate contracting myotubes comprising mature Z-discs. Using a quantitative proteomics approach, we found significant changes in the relative abundance of 387 proteins in myoblastsversusdifferentiated myotubes, reflecting the drastic phenotypic conversion of these cells during myogenesis. Interestingly, EPS of differentiated myotubes to induce Z-disc assembly and maturation resulted in increased levels of proteins involved in ATP synthesis, presumably to fulfill the higher energy demand of contracting myotubes. Because an important role of the Z-disc for signal integration and transduction was recently suggested, its precise phosphorylation landscape further warranted in-depth analysis. We therefore established, by global phosphoproteomics of EPS-treated contracting myotubes, a comprehensive site-resolved protein phosphorylation map of the Z-disc and found that it is a phosphorylation hotspot in skeletal myocytes, underscoring its functions in signaling and disease-related processes. In an illustrative fashion, we analyzed the actin-binding multiadaptor protein filamin C (FLNc), which is essential for Z-disc assembly and maintenance, and found that PKCα phosphorylation at distinct serine residues in its hinge 2 region prevents its cleavage at an adjacent tyrosine residue by calpain 1. Fluorescence recovery after photobleaching experiments indicated that this phosphorylation modulates FLNc dynamics. Moreover, FLNc lacking the cleaved Ig-like domain 24 exhibited remarkably fast kinetics and exceedingly high mobility. Our data set provides research community resource for further identification of kinase-mediated changes in myofibrillar protein interactions, kinetics, and mobility that will greatly advance our understanding of Z-disc dynamics and signaling.
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影响因子：
5.5
作者：
Hartman, Thomas J.;Martin, Jody L.;Russell, Brenda
通讯作者：
Russell, Brenda
DOI：
10.1073/pnas.98.4.1595
发表时间：
2001-02
影响因子：
11.1
作者：
F. Takada;Douglas L. Vander Woude;H. Tong;T. Thompson;S. Watkins;L. Kunkel;A. Beggs
通讯作者：
F. Takada;Douglas L. Vander Woude;H. Tong;T. Thompson;S. Watkins;L. Kunkel;A. Beggs
影响因子：
12.7
作者：
Fürst DO;Goldfarb LG;Kley RA;Vorgerd M;Olivé M;van der Ven PF
通讯作者：
van der Ven PF
DOI：
--
发表时间：
2006
期刊：
The international journal of biochemistry & cell biology
影响因子：
--
作者：
F. Raynaud;Carole Jond-Necand;A. Marcilhac;D. Fürst;Y. Benyamin
通讯作者：
F. Raynaud;Carole Jond-Necand;A. Marcilhac;D. Fürst;Y. Benyamin
将XIN重复蛋白鉴定为Nebulin和Nebulette的SH3结构域的新型配体，并分析其在肌纤维形成和重塑过程中的相互作用。
影响因子：
3.3
作者：
Eulitz S;Sauer F;Pelissier MC;Boisguerin P;Molt S;Schuld J;Orfanos Z;Kley RA;Volkmer R;Wilmanns M;Kirfel G;van der Ven PF;Fürst DO
通讯作者：
Fürst DO