Clausen Lind Andrea, De Castro Gomes Davi, Bisquert Ricardo, Mårtensson Jonas, Sundqvist Martina, Forsman Huamei, Dahlgren Claes, David Florian, Siewers Verena
Department of Life Sciences, Chalmers University of Technology, 412 96 Gothenburg, Sweden.
Department of Genetics, Evolution, Microbiology and Immunology, Institute of Biology, University of Campinas, 13083-862 Campinas, São Paulo, Brazil.
FEMS Yeast Res. 2025 Jan 30;25. doi: 10.1093/femsyr/foaf001.
Yeast-based sensors have shown great applicability for deorphanization of G protein-coupled receptors (GPCRs) and screening of ligands targeting these. A GPCR of great interest is free fatty acid 2 receptor (FFA2R), for which short-chain fatty acids such as propionate and acetate are agonists. FFA2R regulates a wide array of downstream receptor signaling pathways in both adipose tissue and immune cells and has been recognized as a promising therapeutic target, having been implicated in several metabolic and inflammatory diseases. While research aiming to identify ligands recognized by FFA2R for translational applications is ongoing, screening is complicated by the complex regulatory and cell-specific responses mediated by the receptor. To simplify screening towards identification of novel ligands, heterologous platforms are valuable tools that offer efficient identification of ligand activity in the absence of regulatory mechanisms. Here, we present a yeast-based sensor designed to evaluate G protein α i1-mediated FFA2R signaling, with an assay time of 3 h. We verify this platform towards the natural agonists, propionate and acetate, and show applicability towards evaluation of synthetic agonists, antagonists, and allosteric agonists. As such, we believe that the developed yeast strain constitutes a promising screening platform for effective evaluation of ligands acting on FFA2R.
基于酵母的传感器已在G蛋白偶联受体（GPCR）的去孤儿化以及针对这些受体的配体筛选方面显示出巨大的适用性。一种备受关注的GPCR是游离脂肪酸2受体（FFA2R），丙酸酯和乙酸酯等短链脂肪酸是其激动剂。FFA2R在脂肪组织和免疫细胞中调节一系列下游受体信号通路，并且已被认为是一个有前景的治疗靶点，与多种代谢和炎症性疾病有关。虽然旨在鉴定FFA2R识别的用于转化应用的配体的研究正在进行中，但由于该受体介导的复杂调节和细胞特异性反应，筛选工作变得复杂。为了简化鉴定新型配体的筛选过程，异源平台是有价值的工具，可在没有调节机制的情况下有效鉴定配体活性。在此，我们展示了一种基于酵母的传感器，用于评估G蛋白α i1介导的FFA2R信号传导，测定时间为3小时。我们针对天然激动剂丙酸酯和乙酸酯验证了该平台，并展示了其在评估合成激动剂、拮抗剂和变构激动剂方面的适用性。因此，我们认为所开发的酵母菌株构成了一个有前景的筛选平台，可有效评估作用于FFA2R的配体。
