Department of Biomedical Sciences, School of Medicine, University of Missouri-Kansas City, 2411 Holmes Street, Kansas City, MO, 64108, USA.
Department of Anesthesiology, School of Medicine, University of Missouri-Kansas City, 2411 Holmes Street, Kansas City, MO, 64108, USA.
J Mol Neurosci. 2018 Dec;66(4):629-638. doi: 10.1007/s12031-018-1193-0. Epub 2018 Nov 15.
The metabotropic glutamate (mGlu) receptor 5 is a G protein-coupled receptor and is densely expressed in the mammalian brain. Like other glutamate receptors, mGlu5 receptors are tightly regulated by posttranslational modifications such as phosphorylation, although underlying mechanisms are incompletely investigated. In this study, we investigated the role of a prime kinase, extracellular signal-regulated kinase 1 (ERK1), in the phosphorylation and regulation of mGlu5 receptors in vitro and in striatal neurons. We found that recombinant ERK1 proteins directly bound to the C-terminal tail (CT) of mGlu5 receptors in vitro. Endogenous ERK1 also interacted with mGlu5 receptor proteins in adult rat striatal neurons in vivo. The kinase showed the ability to phosphorylate mGlu5 receptors. A serine residue in the distal region of mGlu5 CT was found to be a primary phosphorylation site sensitive to ERK1. In functional studies, we found that pharmacological inhibition of ERK with an inhibitor U0126 reduced the efficacy of mGlu5 receptors in stimulating production of cytoplasmic inositol-1,4,5-triphosphate, a major downstream conventional signaling event, in striatal neurons under normal conditions. These results identify mGlu5 as a new biochemical substrate of ERK1. The kinase can interact with and phosphorylate an intracellular domain of mGlu5 receptors in striatal neurons and thereby control its signaling efficacy.
代谢型谷氨酸(mGlu)受体 5 是一种 G 蛋白偶联受体,在哺乳动物大脑中高度表达。与其他谷氨酸受体一样,mGlu5 受体的翻译后修饰(如磷酸化)受到严格调控,尽管其潜在机制尚未完全研究清楚。在这项研究中,我们研究了原激酶细胞外信号调节激酶 1(ERK1)在体外和纹状体神经元中 mGlu5 受体磷酸化和调节中的作用。我们发现,重组 ERK1 蛋白在体外直接与 mGlu5 受体的 C 端尾部(CT)结合。内源性 ERK1 也与体内成年大鼠纹状体神经元中的 mGlu5 受体蛋白相互作用。该激酶显示出磷酸化 mGlu5 受体的能力。mGlu5 CT 远端区域的一个丝氨酸残基被发现是对 ERK1 敏感的主要磷酸化位点。在功能研究中,我们发现药理学抑制剂 U0126 抑制 ERK 会降低 mGlu5 受体在正常情况下刺激纹状体神经元产生细胞质肌醇 1,4,5-三磷酸的效力,肌醇 1,4,5-三磷酸是主要的下游传统信号事件。这些结果表明 mGlu5 是 ERK1 的一种新的生化底物。该激酶可以与纹状体神经元中的 mGlu5 受体的细胞内结构域相互作用并磷酸化该结构域,从而控制其信号转导效率。