The G protein coupled receptor CXCR4 designed by the QTY code becomes more hydrophilic and retains cell signaling activity

Sci Rep. 2020 Dec 7;10(1):21371. doi: 10.1038/s41598-020-77659-x.

Abstract

G protein-coupled receptors (GPCRs) are vital for diverse biological functions, including vision, smell, and aging. They are involved in a wide range of diseases, and are among the most important targets of medicinal drugs. Tools that facilitate GPCR studies or GPCR-based technologies or therapies are thus critical to develop. Here we report using our QTY (glutamine, threonine, tyrosine) code to systematically replace 29 membrane-facing leucine, isoleucine, valine, and phenylalanine residues in the transmembrane α-helices of the GPCR CXCR4. This variant, CXCR4QTY29, became more hydrophilic, while retaining the ability to bind its ligand CXCL12. When transfected into HEK293 cells, it inserted into the cell membrane, and initiated cellular signaling. This QTY code has the potential to improve GPCR and membrane protein studies by making it possible to design functional hydrophilic receptors. This tool can be applied to diverse α-helical membrane proteins, and may aid in the development of other applications, including clinical therapies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chemokine CXCL12 / chemistry
  • Chemokine CXCL12 / metabolism
  • Chromatography, Affinity
  • Circular Dichroism
  • HEK293 Cells
  • Humans
  • Hydrophobic and Hydrophilic Interactions
  • Membrane Proteins / chemistry*
  • Membrane Proteins / metabolism*
  • Microscopy, Confocal
  • Protein Binding
  • Protein Structure, Secondary
  • Receptors, CXCR4 / chemistry*
  • Receptors, CXCR4 / metabolism*
  • Receptors, G-Protein-Coupled / chemistry
  • Receptors, G-Protein-Coupled / metabolism
  • Signal Transduction / genetics
  • Signal Transduction / physiology

Substances

  • CXCL12 protein, human
  • CXCR4 protein, human
  • Chemokine CXCL12
  • Membrane Proteins
  • Receptors, CXCR4
  • Receptors, G-Protein-Coupled