Department of Physiology and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio, USA.
Department of Medicine, The University of Pittsburgh, Pittsburgh, Pennsylvania, USA.
J Cell Biochem. 2021 Aug;122(8):827-834. doi: 10.1002/jcb.29919. Epub 2021 Apr 13.
G protein-coupled receptors regulate a variety of cellular responses and have been considered as therapeutic targets for human diseases. Lysophosphatidic acid receptor 1 (LPA1) is a receptor for bioactive lysophospholipid, LPA. LPA/LPA1-mediated signaling contributes to inflammatory and fibrotic responses in lung diseases; thus understanding regulation of LPA1 stability is important for modulating LPA/LPA1 signaling. Our previous study has shown that LPA1 is degraded in the Nedd4 like (Nedd4L) E3 ubiquitin ligase-mediated ubiquitin-proteasome system. In the current study, we attempt to identify a peptide that stabilizes LPA1 through disrupting LPA1 association with Nedd4L. LPA treatment induces both endogenous and overexpressed LPA1 degradation, which is attenuated by a proteasome inhibitor, suggesting that LPA1 is degraded in the proteasome. LPA increases phosphorylation of extracellular signal-regulated kinase 1/2 (Erk1/2) and I-κB kinase in lung epithelial cells, and this effect is promoted by overexpression of a peptide (P1) that mimics C-terminal of LPA1. P1, not a control peptide, attenuates LPA-induced LPA1 ubiquitination and degradation, suggesting that P1 stabilizes LPA1. Further, P1 diminishes Nedd4L-mediated degradation of LPA1 and Nedd4L/LPA1 association. In addition to increasing LPA1 signaling, P1 enhances LPA-induced cell migration and gene expression of Elafin, matrix metallopeptidase 1, and serpin family B member 2 in lung epithelial cells. These data suggest that disruption of LPA1 interaction with Nedd4L by P1 increases LPA1 stability and LPA/LPA1 signaling.
G 蛋白偶联受体调节多种细胞反应,被认为是人类疾病的治疗靶点。溶血磷脂酸受体 1(LPA1)是生物活性溶血磷脂酸(LPA)的受体。LPA/LPA1 介导的信号转导有助于肺部疾病中的炎症和纤维化反应;因此,理解 LPA1 稳定性的调节对于调节 LPA/LPA1 信号转导非常重要。我们之前的研究表明,LPA1 在 Nedd4 样(Nedd4L)E3 泛素连接酶介导的泛素-蛋白酶体系统中降解。在本研究中,我们试图通过破坏 LPA1 与 Nedd4L 的结合来鉴定一种稳定 LPA1 的肽。LPA 处理诱导内源性和过表达的 LPA1 降解,该降解被蛋白酶体抑制剂减弱,表明 LPA1 在蛋白酶体中降解。LPA 增加肺上皮细胞中细胞外信号调节激酶 1/2(Erk1/2)和 I-κB 激酶的磷酸化,并且这种效应通过模拟 LPA1 C 末端的肽(P1)的过表达来促进。P1(而非对照肽)减弱 LPA 诱导的 LPA1 泛素化和降解,表明 P1 稳定 LPA1。此外,P1 减弱 Nedd4L 介导的 LPA1 降解和 Nedd4L/LPA1 结合。除了增加 LPA1 信号转导外,P1 还增强 LPA 诱导的肺上皮细胞中 Elafin、基质金属蛋白酶 1 和丝氨酸蛋白酶家族 B 成员 2 的基因表达。这些数据表明,P1 通过破坏 LPA1 与 Nedd4L 的相互作用增加了 LPA1 的稳定性和 LPA/LPA1 信号转导。