Division of Drug Discovery and Safety, Leiden Academic Centre for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands.
Department of Molecular Hematology, Sanquin Research, Plesmalaan 125, 1066 CX Amsterdam, The Netherlands.
J Med Chem. 2023 Aug 24;66(16):11399-11413. doi: 10.1021/acs.jmedchem.3c00854. Epub 2023 Aug 2.
The adenosine A receptor (AAR) is a G protein-coupled receptor (GPCR) that exerts immunomodulatory effects in pathophysiological conditions such as inflammation and cancer. Thus far, studies toward the downstream effects of AAR activation have yielded contradictory results, thereby motivating the need for further investigations. Various chemical and biological tools have been developed for this purpose, ranging from fluorescent ligands to antibodies. Nevertheless, these probes are limited by their reversible mode of binding, relatively large size, and often low specificity. Therefore, in this work, we have developed a clickable and covalent affinity-based probe (AfBP) to target the human AAR. Herein, we show validation of the synthesized AfBP in radioligand displacement, SDS-PAGE, and confocal microscopy experiments as well as utilization of the AfBP for the detection of endogenous AAR expression in flow cytometry experiments. Ultimately, this AfBP will aid future studies toward the expression and function of the AAR in pathologies.
腺苷 A 受体 (AAR) 是一种 G 蛋白偶联受体 (GPCR),在炎症和癌症等病理生理条件下发挥免疫调节作用。到目前为止,对 AAR 激活的下游效应的研究产生了相互矛盾的结果,因此需要进一步的研究。为此目的已经开发了各种化学和生物工具,范围从荧光配体到抗体。然而,这些探针受到其结合的可逆模式、相对较大的尺寸以及通常较低的特异性的限制。因此,在这项工作中,我们开发了一种可点击和基于共价亲和力的探针 (AfBP) 来靶向人类 AAR。在此,我们展示了合成的 AfBP 在放射性配体置换、SDS-PAGE 和共聚焦显微镜实验中的验证,以及在流式细胞术实验中利用 AfBP 检测内源性 AAR 表达的情况。最终,该 AfBP 将有助于未来研究 AAR 在病理学中的表达和功能。