G protein-Coupled Receptors: An Overview of Signaling Mechanisms and Screening Assays

Author(s):  
Duarte Miguel F. Prazeres ◽  
Sofia Aires M. Martins
2019 ◽  
Vol 1456 (1) ◽  
pp. 5-25 ◽  
Author(s):  
Rory K. Morgan ◽  
Garret R. Anderson ◽  
Demet Araç ◽  
Gabriela Aust ◽  
Nariman Balenga ◽  
...  

2014 ◽  
Vol 1333 (1) ◽  
pp. 43-64 ◽  
Author(s):  
Ines Liebscher ◽  
Brian Ackley ◽  
Demet Araç ◽  
Donna M. Ariestanti ◽  
Gabriela Aust ◽  
...  

2009 ◽  
Vol 9 (2) ◽  
pp. 242-250 ◽  
Author(s):  
Yonglin Wang ◽  
Aining Li ◽  
Xiaoli Wang ◽  
Xin Zhang ◽  
Wei Zhao ◽  
...  

ABSTRACT G protein-coupled receptors (GPCRs) represent a large receptor family involved in a broad spectrum of cell signaling. To understand signaling mechanisms mediated by GPCRs in Phytophthora sojae, we identified and characterized the PsGPR11 gene, which encodes a putative seven-transmembrane GPCR. An expression analysis revealed that PsGPR11 was differentially expressed during asexual development. The highest expression level occurred in zoospores and was upregulated during early infection. PsGPR11-deficienct transformants were obtained by gene silencing strategies. Silenced transformants exhibited no differences in hyphal growth or morphology, sporangium production or size, or mating behavior. However, the release of zoospores from sporangia was severely impaired in the silenced transformants, and about 50% of the sporangia did not completely release their zoospores. Zoospore encystment and germination were also impaired, and zoospores of the transformants lost their pathogenicity to soybean. In addition, no interaction was observed between PsGPR11 and PsGPA1 with a conventional yeast two-hybrid assay, and the transcriptional levels of some genes which were identified as being negatively regulated by PsGPA1 were not clearly altered in PsGPR11-silenced mutants. These results suggest that PsGPR11-mediated signaling controls P. sojae zoospore development and virulence through the pathways independent of G protein.


2003 ◽  
Vol 8 (1) ◽  
pp. 39-49 ◽  
Author(s):  
Andrew M.F. Liu ◽  
Maurice K.C. Ho ◽  
Cecilia S.S. Wong ◽  
Jasmine H.P. Chan ◽  
Anson H.M. Pau ◽  
...  

G protein—coupled receptors (GPCRs) represent a class of important therapeutic targets for drug discovery. The integration of GPCRs into contemporary high-throughput functional assays is critically dependent on the presence of appropriate G proteins. Given that different GPCRs can discriminate against distinct G proteins, a universal G protein adapter is extremely desirable. In this report, the authors evaluated two highly promiscuous Gα16/z chimeras, 16z25 and 16z44, for their ability to translate GPCR activation into Ca2+ mobilization using the fluorescence imaging plate reader (FLIPR) and aequorin. A panel of 24 Gs- or Gi-coupled receptors was examined for their functional association with the Gα16/z chimeras. Although most of the GPCRs tested were incapable of inducing Ca2+ mobilization upon their activation by specific agonists, the introduction of 16z25 or 16z44 allowed all of these GPCRs to mediate agonist-induced Ca 2+ mobilization. In contrast, only 16 of the GPCRs tested were capable of using Gα16 to mobilize intracellular Ca2+. Analysis of dose-response curves obtained with the δ-opioid, dopamine D1 , and Xenopus melatonin Mel1c receptors revealed that the Gα 16/z chimeras possess better sensitivity than Gα16 in both the FLIPR and aequorin assays. Collectively, these studies help to validate the promiscuity of the Gα16/z chimeras as well as their application in contemporary drug-screening assays that are based on ligand-induced Ca 2+ mobilization. ( Journal of Biomolecular Screening 2003:39-49)


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