Comparing Native Crystal Structures and AlphaFold2 Predicted Water-Soluble G Protein-Coupled Receptor QTY Variants

Accurate predictions of 3-dimensional protein structures by AlphaFold2 is a game-changer for biology, especially for structural biology. Here we present the studies of several native chemokine receptors including CCR5, CCR9, CXCR2 and CXCR4 determined by X-ray crystallography, and their water-soluble QTY counter parts predicted by AlphaFold2. In the native structures, there are hydrophobic amino acids leucine (L), isoleucine (I), valine (V) and phenylalanine (F) in the transmembrane helices. These hydrophobic amino acids are systematically replaced by hydrophilic amino acids glutamine (Q), threonine (T), and tyrosine (Y). Thus, the QTY variants become water-soluble. We also present the superimposed structures of native CCR10, CXCR5, CXCR7 and an olfactory receptor OR1D2 and their water-soluble QTY variants. Since the CryoEM structural determinations for the QTY variants of CCR10(QTY) and OR1D2(QTY) are in progress, it will be of interest to compare them when the structures become available. The superimposed structures show remarkable similarity within RMSD 1 angstrom-2 angstrom despite significant sequence differences (~26%-~33%). We also show the differences of hydrophobicity patches between the native GPCR and their QTY variants. Our study provides insight into the subtle differences between the hydrophobic helices and hydrophilic helices, and may further stimulate designs of water-soluble membrane proteins and other aggregated proteins.

Automated summary

The accurate predictions of protein structures by AlphaFold2 have revolutionized biology, particularly structural biology. This study compared native chemokine receptor structures with their water-soluble QTY variants, showing high structural similarity despite significant sequence differences. Insights gained from this study may facilitate the design of water-soluble membrane proteins and other aggregated proteins.