In Vivo Phosphoproteomics Analysis Reveals the Cardiac Targets of β-Adrenergic Receptor Signaling
In Vivo Phosphoproteomics Analysis Reveals the Cardiac Targets of β-Adrenergic Receptor Signaling
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DOI：
10.1126/scisignal.2003506
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发表时间：
2013-06-04
影响因子：
7.3
通讯作者：
Olsen, Jesper V.
中科院分区：
文献类型：
--
作者：
Lundby, Alicia;Andersen, Martin N.;Olsen, Jesper V.
β-阻滞剂广泛用于预防心律失常和通过抑制β-肾上腺素能受体（β AR）从而降低收缩性和心率来治疗高血压。β AR启动磷酸化依赖的信号级联，但只有少数靶蛋白是已知的。我们使用定量体内磷酸化蛋白质组学来鉴定小鼠心脏中670个位点特异性磷酸化变化，以响应特定β AR激动剂的急性治疗。邻近调节磷酸化位点的残基表现出序列特异性偏好（R-X-X-pS/T），序列基序和相互作用网络的综合分析表明，AMPK激酶（腺苷5 '-单磷酸活化蛋白激酶）、Akt和mTOR（雷帕霉素的哺乳动物靶标）介导β AR信号传导，除了由PKA（环腺苷一磷酸依赖性蛋白激酶）和CaMKII（钙/钙调蛋白依赖性蛋白激酶II型）介导的成熟途径之外。我们发现了6个离子通道和转运蛋白上的磷酸化位点的特异性调节，这些离子通道和转运蛋白介导了较高心率下离子通量的增加，我们发现其中一个，钾通道K（V）7.1的Ser（92）的磷酸化增加了电流幅度。我们的数据集代表了一个定量分析的磷酸化蛋白质在体内调节后，刺激的七跨膜受体，我们的研究结果揭示了以前未知的磷酸化位点，调节心肌收缩力，这表明新的潜在目标，用于治疗心脏病和高血压。
beta-Blockers are widely used to prevent cardiac arrhythmias and to treat hypertension by inhibiting beta-adrenergic receptors (beta ARs) and thus decreasing contractility and heart rate. beta ARs initiate phosphorylation-dependent signaling cascades, but only a small number of the target proteins are known. We used quantitative in vivo phosphoproteomics to identify 670 site-specific phosphorylation changes in murine hearts in response to acute treatment with specific beta AR agonists. The residues adjacent to the regulated phosphorylation sites exhibited a sequence-specific preference (R-X-X-pS/T), and integrative analysis of sequence motifs and interaction networks suggested that the kinases AMPK (adenosine 5'-monophosphate-activated protein kinase), Akt, and mTOR (mammalian target of rapamycin) mediate beta AR signaling, in addition to the well-established pathways mediated by PKA (cyclic adenosine monophosphate-dependent protein kinase) and CaMKII (calcium/calmodulin-dependent protein kinase type II). We found specific regulation of phosphorylation sites on six ion channels and transporters that mediate increased ion fluxes at higher heart rates, and we showed that phosphorylation of one of these, Ser(92) of the potassium channel K(V)7.1, increased current amplitude. Our data set represents a quantitative analysis of phosphorylated proteins regulated in vivo upon stimulation of seven-transmembrane receptors, and our findings reveal previously unknown phosphorylation sites that regulate myocardial contractility, suggesting new potential targets for the treatment of heart disease and hypertension.