Identification of novel species-selective agonists of the G-protein-coupled receptor GPR35 that promote recruitment of β-arrestin-2 and activate Gα13
Identification of novel species-selective agonists of the G-protein-coupled receptor GPR35 that promote recruitment of β-arrestin-2 and activate Gα13
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DOI：
10.1042/bj20101287
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发表时间：
2010-12-15
影响因子：
4.1
通讯作者：
Milligan, Graeme
中科院分区：
文献类型：
--
作者：
Jenkins, Laura;Brea, Jose;Milligan, Graeme
G蛋白偶联受体GPR35的特性不佳，已被认为是一个潜在的探索性靶点，用于治疗代谢紊乱和高血压。它也被证明在免疫调节中发挥重要作用。验证这些概念和进一步研究这种受体的作用的主要障碍是缺乏与GPR35相互作用的药理学工具。使用受体-β-arrestin-2相互作用测定与人类和大鼠的GPR35的直系同源物，我们确定了一些具有激动剂活性的化合物。这些包括先前描述的配体扎普司特。尽管许多活性化合物，包括克罗维酸二钠和双香豆素，在GPR35的两种直系同源物上显示出相似的效力，但许多配体，包括双羟萘酸和尼氟灭酸，仅在人GPR35上具有可检测的活性，而其他配体，包括扎普司特和木犀草素，对大鼠直系同源物具有显著的选择性。先前的研究已经证明G（α 13）被GPR35激活。采用嵌合Gpal-G（α 13）G蛋白的基于酵母的测定证实，在β-抑制蛋白-2相互作用测定中，所有对人GPR35有活性的化合物也能够通过G（α 13）促进细胞生长。这些配体中的每一种也以GPR35依赖性方式促进[S-35] GTP [S]（鸟苷5 ′-[γ-[S-35]硫代]三磷酸）与表位标记形式的G13的结合。在这些研究中确定的配体将是有用的询问GPR35的生物学作用，但在这个受体的配体的物种选择性的升值将是至关重要的正确属性功能。
The poorly characterized G-protein-coupled receptor GPR35 has been suggested as a potential exploratory target for the treatment of both metabolic disorders and hypertension. It has also been indicated to play an important role in immune modulation. A major impediment to validation of these concepts and further study of the role of this receptor has been a paucity of pharmacological tools that interact with GPR35. Using a receptor-beta-arrestin-2 interaction assay with both human and rat orthologues of GPR35, we identified a number of compounds possessing agonist activity. These included the previously described ligand zaprinast. Although a number of active compounds, including cromolyn disodium and dicumarol, displayed similar potency at both orthologues of GPR35, a number of ligands, including pamoate and niflumic acid, had detectable activity only at human GPR35 whereas others, including zaprinast and luteolin, were markedly selective for the rat orthologue. Previous studies have demonstrated activation of G(alpha 13) by GPR35. A Saccharomyces cerevisiae-based assay employing a chimaeric Gpal-G(alpha 13) G-protein confirmed that all of the compounds active at human GPR35 in the beta-arrestin-2 interaction assay were also able to promote cell growth via G(alpha 13). Each of these ligands also promoted binding of [S-35]GTP[S] (guanosine 5'-[gamma-[S-35]thio]triphosphate) to an epitope-tagged form of G,3 in a GPR35-dependent manner. The ligands identified in these studies will be useful in interrogating the biological actions of GPR35, but appreciation of the species selectivity of ligands at this receptor will be vital to correctly attribute function.