QTY code enables design of detergent-free chemokine receptors that retain ligand-binding activities

Structure and function studies of membrane proteins, particularly G protein-coupled receptors and multipass transmembrane proteins, require detergents. We have devised a simple tool, the QTY code (glutamine, threonine, and tyrosine), for designing hydrophobic domains to become water soluble without detergents. Here we report using the QTY code to systematically replace the hydro-phobic amino acids leucine, valine, isoleucine, and phenylalanine in the seven transmembrane alpha-helices of CCR5, CXCR4, CCR10, and CXCR7. We show that QTY code-designed chemokine receptor variants retain their thermostabilities, alpha-helical structures, and ligand-binding activities in buffer and 50% human serum. CCR5(QTY), CXCR4(QTY), and CXCR7(QTY) also bind to HIV coat protein gp41-120. Despite substantial transmembrane domain changes, the detergent-free QTY variants maintain stable structures and retain their ligand-binding activities. We believe the QTY code will be use-ful for designing water-soluble variants of membrane proteins and other water-insoluble aggregated proteins.