Optimization of Angiotensin II Receptor Type 1 Production for Biophysical Studies

The class of G protein-coupled receptors (GPCRs) represents a ubiquitous family of membrane proteins that selectively interact with a variety of signaling molecules to modulate cellular biochemical and physiological processes. Consequently, the structural and functional investigation of GPCRs is of paramount importance for both fundamental and applied scientific research. However, challenges arise in their recombinant expression, as well as subsequent extraction and purification, due to their inherent thermal instability and low yield. This necessitates the optimization of expression conditions and protocols for solubilization and purification. In this study, we focused on optimizing the expression, solubilization, and purification processes for the angiotensin II type 1 receptor (AT1), which plays a critical role in the physiology of the cardiovascular system. Several critical factors affecting receptor expression levels, yield, and monomericity were identified, facilitating an enhancement in the quality of the produced protein.