Food Chemistry, Department of Chemistry and Pharmacy, Friedrich-Alexander-Universität Erlangen-Nürnberg (FAU), Nikolaus-Fiebiger-Str. 10, 91058, Erlangen, Germany.
Princess Margaret Cancer Centre, University Health Network, 101 College Street, Toronto, ON, Canada.
Sci Rep. 2021 Feb 10;11(1):3501. doi: 10.1038/s41598-021-83038-x.
The development of functionally selective or biased ligands is a promising approach towards drugs with less side effects. Biased ligands for G protein-coupled receptors can selectively induce G protein activation or β-arrestin recruitment. The consequences of this selective action on cellular functions, however, are not fully understood. Here, we investigated the impact of five biased and balanced dopamine D receptor agonists and antagonists on the global protein expression in HEK293T cells by untargeted nanoscale liquid chromatography-tandem mass spectrometry. The proteome analysis detected 5290 protein groups. Hierarchical clustering and principal component analysis based on the expression levels of 1462 differential proteins led to a separation of antagonists and balanced agonist from the control treatment, while the biased ligands demonstrated larger similarities to the control. Functional analysis of affected proteins revealed that the antagonists haloperidol and sulpiride regulated exocytosis and peroxisome function. The balanced agonist quinpirole, but not the functionally selective agonists induced a downregulation of proteins involved in synaptic signaling. The β-arrestin-preferring agonist BM138, however, regulated several proteins related to neuron function and the dopamine receptor-mediated signaling pathway itself. The G protein-selective partial agonist MS308 influenced rather broad functional terms such as DNA processing and mitochondrial translation.
开发具有功能选择性或偏向性的配体是一种很有前途的方法，可以减少药物的副作用。G 蛋白偶联受体的偏向性配体可以选择性地诱导 G 蛋白激活或β-arrestin 募集。然而，这种选择性作用对细胞功能的影响尚不完全清楚。在这里，我们通过非靶向纳米液相色谱-串联质谱法研究了五种偏向性和平衡的多巴胺 D 受体激动剂和拮抗剂对 HEK293T 细胞中全局蛋白质表达的影响。蛋白质组分析检测到 5290 个蛋白质组。基于 1462 个差异蛋白表达水平的层次聚类和主成分分析导致拮抗剂和平衡激动剂与对照处理分离，而偏向性配体与对照更为相似。受影响蛋白的功能分析表明，拮抗剂氟哌啶醇和舒必利调节细胞外排和过氧化物酶体功能。平衡激动剂喹吡罗尔，但不是功能选择性激动剂，下调了参与突触信号转导的蛋白。然而，β-arrestin 偏好性激动剂 BM138 调节与神经元功能和多巴胺受体介导的信号通路本身相关的几种蛋白质。G 蛋白选择性部分激动剂 MS308 影响的功能术语则更为广泛，如 DNA 处理和线粒体翻译。
