Identification of Phosphorylation and Other Post-Translational Modifications in the Central C4C5 Domains of Murine Cardiac Myosin Binding Protein C.
Identification of Phosphorylation and Other Post-Translational Modifications in the Central C4C5 Domains of Murine Cardiac Myosin Binding Protein C.
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DOI：
10.1021/acsomega.2c00799
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发表时间：
2022-04-26
期刊：
影响因子：
4.1
通讯作者：
Stelzer, Julian E.
中科院分区：
文献类型：
--
作者：
Doh, Chang Yoon;Dominic, Katherine L.;Swanberg, Caitlin E.;Bharambe, Nikhil;Willard, Belinda B.;Li, Ling;Ramachandran, Rajesh;Stelzer, Julian E.
心肌肌球蛋白结合蛋白C(CMyBPC)是一种调节肌球蛋白跨桥行为和心肌收缩能力的重要多域蛋白。CMyBPC主要受其N-末端M-结构域内残基的磷酸化调控。然而，对中央C4C5结构域的磷酸化或其他翻译后修饰(PTM)情况知之甚少。在这项研究中，利用表达带有不可磷酸化丝氨酸的cMyBPC(S)到丙氨酸取代的小鼠模型，在体内证实了M结构域之外的磷酸化的存在。将纯化的重组小鼠C4C5结构域与13种不同的激酶孵育，并从6种最强的激酶中选取样品进行质谱分析。共发现26个独特的磷酸化多肽，代表13个不同的磷酸化位点，其中包括10个新的位点。平行反应监测和随后的诱变实验表明，S690位点(UniProtKB O70468)是PKA和PKG1的主要靶点。我们还报告了文献中没有描述的6个乙酰化和7个泛素化位点。这些PTM证明了cMyBPC的中心结构域在心脏健康和疾病中具有额外调控层的可能性和潜在的重要性。数据可通过标识符为PXD031262的ProteomeXchange获得。
Cardiac myosin binding protein C (cMyBPC) is a critical multidomain protein that modulates myosin cross bridge behavior and cardiac contractility. cMyBPC is principally regulated by phosphorylation of the residues within the M-domain of its N-terminus. However, not much is known about the phosphorylation or other post-translational modification (PTM) landscape of the central C4C5 domains. In this study, the presence of phosphorylation outside the M-domain was confirmed in vivo using mouse models expressing cMyBPC with nonphosphorylatable serine (S) to alanine substitutions. Purified recombinant mouse C4C5 domain constructs were incubated with 13 different kinases, and samples from the 6 strongest kinases were chosen for mass spectrometry analysis. A total of 26 unique phosphorylated peptides were found, representing 13 different phosphorylation sites including 10 novel sites. Parallel reaction monitoring and subsequent mutagenesis experiments revealed that the S690 site (UniProtKB O70468) was the predominant target of PKA and PKG1. We also report 6 acetylation and 7 ubiquitination sites not previously described in the literature. These PTMs demonstrate the possibility of additional layers of regulation and potential importance of the central domains of cMyBPC in cardiac health and disease. Data are available via ProteomeXchange with identifier PXD031262.
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影响因子：
2.9
作者：
Belknap, Betty;Harris, Samantha P.;White, Howard D.
通讯作者：
White, Howard D.
DOI：
10.1016/j.bbadis.2019.01.007
发表时间：
2019-03-01
影响因子：
6.2
作者：
Doh, Chang Yoon;Li, Jiayang;Stelzer, Julian E.
通讯作者：
Stelzer, Julian E.
影响因子：
37.8
作者：
Decker, RS;Decker, ML;Winegrad, S
通讯作者：
Winegrad, S
DOI：
10.1073/pnas.1521281113
发表时间：
2016-03-22
影响因子：
11.1
作者：
Colson, Brett A.;Thompson, Andrew R.;Thomas, David D.
通讯作者：
Thomas, David D.
影响因子：
5.5
作者：
Gresham, Kenneth S.;Stelzer, Julian E.
通讯作者：
Stelzer, Julian E.