Abstract
Overexpression of biologically functional GPCRs and homogeneous purified protein solutions are required to enable structural studies and protein-based biophysical assay development. Iterative and time-consuming optimization cycles of protein engineering, expression, and purification are often needed to achieve the desired protein quantity and quality. Here, we describe the reconstitution of GPCRs in virus-like particles (VLPs) and their use in biophysical assays to characterize protein yield, stability, and small molecule ligand binding. This approach prevents the need for time-consuming detergent solubilization and protein purification during recombinant GPCR protein optimization.
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Acknowledgments
The authors would like to thank Esther Peluso for generating the figures; Alex Capovilla, Jennifer Nguyen, and Leonard Kaljevic for critical review of the manuscript.
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Aertgeerts, K., Ho, T.T., Yan, Y.G. (2022). Optimization of Recombinant GPCR Proteins for Biophysical and Structural Studies Using Virus-like Particles. In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology, vol 2507. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2368-8_17
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DOI: https://doi.org/10.1007/978-1-0716-2368-8_17
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