Structural and functional characterization of membrane proteins (MPs) requires extraction from the native lipid bilayer, which generally necessitates the formation of water-soluble MPs structures or complexes suitable for biochemical assays. These structures are often detergent micelles that form protein-detergent complexes (PDCs), due to detergents being straightforward
in implementation and providing homogenous yields. However, obtaining physiologically relevant PDCs remains a challenge, because most detergents do not keep the native structure/function of the protein intact, jeopardizing its stability, and disrupting the proton pressure and electric gradient even after reincorporation in artificial lipid membranes. To achieve native-like
physiology (i.e. to maintain native protein-associated lipids and structurally relevant ligands or cofactors), it is essential to choose the right extraction parameters. Nevertheless, the field of membrane biochemistry is still missing a holistic, interdependent understanding of these systems, too often neglecting the evident, key role that detergents play. In this review, we aim to obtain a more fundamental understanding of detergent-solubilized membrane proteins by pinpointing the most relevant assay parameters (i.e. the most influential on the biochemical properties) involving MPs.