Optimization of Recombinant GPCR Proteins for Biophysical and Structural Studies Using Virus-like Particles.
Methods Mol Biol
; 2507: 327-336, 2022.
Article
em En
| MEDLINE
| ID: mdl-35773590
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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1
Base de dados:
MEDLINE
Assunto principal:
Receptores Acoplados a Proteínas G
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
Estados Unidos