Phosphorylation bar-coding of free fatty acid receptor 2 is generated in a tissue-specific manner.
Phosphorylation bar-coding of free fatty acid receptor 2 is generated in a tissue-specific manner.
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DOI：
10.7554/elife.91861
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发表时间：
2023-12-12
期刊：
影响因子：
7.7
通讯作者：
Milligan G
中科院分区：
文献类型：
--
作者：
Barki N;Jenkins L;Marsango S;Dedeo D;Bolognini D;Dwomoh L;Abdelmalik AM;Nilsen M;Stoffels M;Nagel F;Schulz S;Tobin AB;Milligan G
游离脂肪酸受体 2 (FFAR2) 由短链脂肪酸激活并广泛表达，包括在白色脂肪细胞以及各种免疫和肠内分泌细胞中。使用野生型人 FFAR2 和仅由设计药物 (DREADD) 变体激活的设计受体，我们探索了该受体在异源细胞系和表达人 FFAR2 或 FFAR2-DREADD 的转基因敲入小鼠系的组织中的激活和磷酸化概况。靶向 pSer296/pSer297 或 pThr306/pThr310 的 FFAR2 磷酸位点特异性抗血清提供了组成型和激动剂介导的磷酸化的敏感生物标志物，以及原位可视化激动剂激活受体的有效方法。在白色脂肪组织中，残基 Ser296/Ser297 的磷酸化在激动剂激活后增强，而 Thr306/Thr310 没有磷酸化。相比之下，在派尔氏集结的免疫细胞中，Thr306/Thr310 以严格激动剂依赖的方式磷酸化，而在结肠的肠内分泌细胞中，Ser296/Ser297 和 Thr306/Thr310 的磷酸化程度很差。迄今为止，磷酸化条形码的概念集中于不同激动剂促进不同受体磷酸化模式的潜力。在这里，我们证明这种情况发生在不同病理生理相关靶组织中的相同激动剂-受体配对中。这可能解释了为什么单个 G 蛋白偶联受体可以以组织特异性方式产生不同的功能结果。
Free fatty acid receptor 2 (FFAR2) is activated by short-chain fatty acids and expressed widely, including in white adipocytes and various immune and enteroendocrine cells. Using both wild-type human FFAR2 and a designer receptor exclusively activated by designer drug (DREADD) variant we explored the activation and phosphorylation profile of the receptor, both in heterologous cell lines and in tissues from transgenic knock-in mouse lines expressing either human FFAR2 or the FFAR2-DREADD. FFAR2 phospho-site-specific antisera targeting either pSer296/pSer297 or pThr306/pThr310 provided sensitive biomarkers of both constitutive and agonist-mediated phosphorylation as well as an effective means to visualise agonist-activated receptors in situ. In white adipose tissue, phosphorylation of residues Ser296/Ser297 was enhanced upon agonist activation whilst Thr306/Thr310 did not become phosphorylated. By contrast, in immune cells from Peyer’s patches Thr306/Thr310 become phosphorylated in a strictly agonist-dependent fashion whilst in enteroendocrine cells of the colon both Ser296/Ser297 and Thr306/Thr310 were poorly phosphorylated. The concept of phosphorylation bar-coding has centred to date on the potential for different agonists to promote distinct receptor phosphorylation patterns. Here, we demonstrate that this occurs for the same agonist-receptor pairing in different patho-physiologically relevant target tissues. This may underpin why a single G protein-coupled receptor can generate different functional outcomes in a tissue-specific manner.
影响因子：
4.6
作者：
Mann A;Keen AC;Mark H;Dasgupta P;Javitch JA;Canals M;Schulz S;Robert Lane J
通讯作者：
Robert Lane J
使用磷酸位点特异性 GPCR 抗体对阿片类药物和大麻素药物作用进行原位可视化。
影响因子：
5.9
作者：
Fritzwanker, Sebastian;Nagel, Falko;Kliewer, Andrea;Stammer, Viviane;Schulz, Stefan
通讯作者：
Schulz, Stefan