Proteomics Analysis of Cellular BRS3 Receptor Activation Reveals Potential Mechanism for Signal Transduction and Cell Proliferation
Proteomics Analysis of Cellular BRS3 Receptor Activation Reveals Potential Mechanism for Signal Transduction and Cell Proliferation
复制标题
细胞 BRS3 受体激活的蛋白质组学分析揭示了信号转导和细胞增殖的潜在机制。
DOI：
10.1021/acs.jproteome.9b00760
复制
发表时间：
2020-04-03
影响因子：
4.4
通讯作者：
Xiao, Hua
中科院分区：
文献类型：
--
作者：
Dong, Lijie;Zhang, Baohui;Xiao, Hua
蛙皮素样受体3(Brs3)是一种孤儿G蛋白偶联受体，在我们的生物系统中发挥着重要的作用，但其确切的作用机制却鲜为人知。为了深入了解Brs3激活的生物学效应，我们利用定量蛋白质组学方法研究了Brs3配体刺激过程中的动态蛋白质谱。在Brs3替代激动剂刺激后的不同时间点，利用纳米LC-MS/MS分析了Brs3过表达的HEK 293细胞Brs3(HEK 293-Brs3)的蛋白质谱，共鉴定和定量了1593个细胞蛋白质，其中146个蛋白质在多个时间点失调，319个蛋白质在一个时间点发生变化。数据分析表明，Brs3的激活可以调节细胞的死亡、存活和蛋白质的合成，特别是mRNA的翻译。揭示了Brs3信号转导的关键信号通路。特别是，我们发现的21个蛋白质与雷帕霉素(MTOR)信号通路有关。通过监测其在Brs3激活时的指示性靶点，进一步证实了mTOR的促进作用。雷帕霉素抑制mTOR后，细胞增殖明显逆转。我们的蛋白质组学数据共同证明，Brs3的激活将导致信号转导的级联反应，促进细胞增殖。制定的战略可能被用来发现其他GPCRs的作用，并增进我们对其未知功能的理解。
Bombesin-like receptor 3 (BRS3), an orphan G protein-coupled receptor (GPCR), plays important roles in our biological system while the exact mechanisms behind are less known. To get insights of biological effects upon BRS3 activation, we utilized quantitative proteomics approach to explore the dynamic protein profiling during the stimulation by its ligand. At different time points after stimulation with BRS3 surrogate agonist, the protein profiling in BRS3 overexpressed HEK 293 cells BRS3 (HEK 293-BRS3) was analyzed by nano-LC-MS/MS. In total, 1593 cellular proteins were confidently identified and quantified, including 146 proteins dysregulated at multiple time points and 319 proteins only altered at one time point. Data analysis indicated that BRS3 activation could regulate cell death, survival, and protein synthesis, particularly mRNA translation. Key signaling pathways were revealed for BRS3 signal transduction. In particular, 21 of our identified proteins involve in rapamycin (mTOR) signaling pathway. The promotion of mTOR was further confirmed through monitoring its indicative targets upon BRS3 activation. Upon the inhibition of mTOR by rapamycin, cell proliferation was dramatically reversed. Our proteomics data collectively demonstrate that BRS3 activation will lead to cascades of signal transduction and promote cell proliferation. The developed strategy might be utilized to discover the roles of other GPCRs and improve our understanding of their unknown functions.