A Novel Allosteric Activator of Free Fatty Acid 2 Receptor Displays Unique Gi-functional Bias.
A Novel Allosteric Activator of Free Fatty Acid 2 Receptor Displays Unique Gi-functional Bias.
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DOI：
10.1074/jbc.m116.736157
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发表时间：
2016-09-02
期刊：
影响因子：
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通讯作者：
Milligan G
中科院分区：
文献类型：
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作者：
Bolognini D;Moss CE;Nilsson K;Petersson AU;Donnelly I;Sergeev E;König GM;Kostenis E;Kurowska-Stolarska M;Miller A;Dekker N;Tobin AB;Milligan G
短链脂肪酸受体FFA2能够通过Gi-和Gq/ g11促进的途径刺激信号传导。这些途径被认为控制着不同的生理终点，但缺乏适合测试这一假设的FFA2受体配体。在此，我们对AZ1729进行了表征，AZ1729是一种新型的FFA2调节剂，作为一种直接变构激动剂和一种正变构调节剂，在gi介导的途径中增加内源性短链脂肪酸丙酸的活性，而不是在Gq/G11转导的途径中。将AZ1729与Gi和Gq/G11家族G蛋白的直接抑制剂联合使用表明，尽管在表达ffa2的293细胞中，两者都参与丙酸介导的磷酸化- erk1 /2 MAP激酶信号的调节，但Gq/G11介导的途径占主导地位。我们通过使用AZ1729来扩展这些研究，剖析生理FFA2信号通路。AZ1729能够作用于FFA2受体，抑制β-肾上腺素受体激动剂促进的小鼠原代脂肪细胞的脂肪分解，并促进分离的人中性粒细胞的趋化性，证实了这些是由Gi信号介导的FFA2过程，然而，与Gq/G11抑制剂FR900359的阻断一致，AZ1729无法模仿或调节丙酸介导的GLP-1从小鼠结肠制剂中的释放，这一生理反应被定义为通过激活Gq/G11转导的终点。
The short chain fatty acid receptor FFA2 is able to stimulate signaling via both Gi- and Gq/G11-promoted pathways. These pathways are believed to control distinct physiological end points but FFA2 receptor ligands appropriate to test this hypothesis have been lacking. Herein, we characterize AZ1729, a novel FFA2 regulator that acts as a direct allosteric agonist and as a positive allosteric modulator, increasing the activity of the endogenously produced short chain fatty acid propionate in Gi-mediated pathways, but not at those transduced by Gq/G11. Using AZ1729 in combination with direct inhibitors of Gi and Gq/G11 family G proteins demonstrated that although both arms contribute to propionate-mediated regulation of phospho-ERK1/2 MAP kinase signaling in FFA2-expressing 293 cells, the Gq/G11-mediated pathway is predominant. We extend these studies by employing AZ1729 to dissect physiological FFA2 signaling pathways. The capacity of AZ1729 to act at FFA2 receptors to inhibit β-adrenoreceptor agonist-promoted lipolysis in primary mouse adipocytes and to promote chemotaxis of isolated human neutrophils confirmed these as FFA2 processes mediated by Gi signaling, whereas, in concert with blockade by the Gq/G11 inhibitor FR900359, the inability of AZ1729 to mimic or regulate propionate-mediated release of GLP-1 from mouse colonic preparations defined this physiological response as an end point transduced via activation of Gq/G11.