GPCR expression and activation in yeast
Perez, Francis E.
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G-protein coupled receptors are cell surface receptors that play a critical role in cell signaling. There is shortage of information about their structures and about the large amount of stimuli that activate them. In our study we expressed GPCRs in yeast to study their interaction with their corresponding ligands. Yeast has similar pathways of GPCR interaction with that of the mammalian pathway and is therefore a useful system for the study. In particular, we developed a yeast-based screening system to study GPCR interaction with different stimuli. Using CXCR4 and SDF-1 as our model system, we expressed CXCR4 on the yeast membrane and test for functionality by SDF-1 activation. CXCR4 and SDF-1 play roles in many diseases, such as cancer and HIV-1, making it important to understand their interaction. We tested for CXCR4 expression by flow cytometry and used beta-galactosidase assay, and a histidine independence growth assay to test for function.