Promega Corporation, 2800 Woods Hollow Road, Madison, WI, 53711, USA.
Promega Biosciences, 277 Granada Drive, San Luis Obispo, CA, 93401, USA.
Sci Rep. 2024 Feb 23;14(1):4440. doi: 10.1038/s41598-024-55038-0.
3',5'-Cyclic adenosine monophosphate (cAMP), the first identified second messenger, is implicated in diverse cellular processes involving cellular metabolism, cell proliferation and differentiation, apoptosis, and gene expression. cAMP is synthesized by adenylyl cyclase (AC), which converts ATP to cAMP upon activation of G-protein coupled receptors (GPCRs) in most cases and hydrolyzed by cyclic nucleotide phosphodiesterases (PDEs) to 5'-AMP. Dysregulation of cAMP signaling is implicated in a wide range of pathophysiological conditions such as cardiovascular diseases, neurodegenerative and behavioral disorders, cancers, diabetes, obesity, cataracts, and others. Therefore, cAMP targeted therapies have been and are still undergoing intense investigation for the treatment of these and other diseases. This highlights the need for developing assays to detect and monitor cAMP levels. In this study, we show cAMP Lumit assay as a highly specific homogeneous bioluminescent assay suitable for high throughput screenings with a large assay window and a wide dynamic range for cAMP detection. We believe that this assay will aid and simplify drug discovery screening efforts for cAMP signaling targeted therapies.
3'，5'-环磷酸腺苷（cAMP），第一种被鉴定的第二信使，参与涉及细胞代谢、细胞增殖和分化、细胞凋亡和基因表达的多种细胞过程。cAMP 是由腺苷酸环化酶（AC）合成的，在大多数情况下，当 G 蛋白偶联受体（GPCR）被激活时，AC 将 ATP 转化为 cAMP，而 cAMP 则被环核苷酸磷酸二酯酶（PDEs）水解为 5'-AMP。cAMP 信号转导的失调与广泛的病理生理状况有关，如心血管疾病、神经退行性和行为障碍、癌症、糖尿病、肥胖症、白内障等。因此，针对 cAMP 的靶向治疗方法一直在进行，并仍在针对这些和其他疾病进行深入研究。这突出表明需要开发检测和监测 cAMP 水平的方法。在本研究中，我们展示了 cAMP Lumit 测定法作为一种高度特异性的均相生物发光测定法，适用于具有大测定窗口和宽动态范围的高通量筛选，用于检测 cAMP。我们相信，该测定法将有助于简化针对 cAMP 信号转导的靶向治疗药物发现筛选工作。
