Towards high throughput GPCR crystallography: In Meso soaking of Adenosine A 2A Receptor crystals

Sci Rep. 2018 Jan 8;8(1):41. doi: 10.1038/s41598-017-18570-w.

Abstract

Here we report an efficient method to generate multiple co-structures of the A2A G protein-coupled receptor (GPCR) with small-molecules from a single preparation of a thermostabilised receptor crystallised in Lipidic Cubic Phase (LCP). Receptor crystallisation is achieved following purification using a low affinity "carrier" ligand (theophylline) and crystals are then soaked in solutions containing the desired (higher affinity) compounds. Complete datasets to high resolution can then be collected from single crystals and seven structures are reported here of which three are novel. The method significantly improves structural throughput for ligand screening using stabilised GPCRs, thereby actively driving Structure-Based Drug Discovery (SBDD).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Crystallography, X-Ray
  • Humans
  • Ligands
  • Models, Molecular
  • Molecular Conformation
  • Protein Unfolding
  • Receptor, Adenosine A2A / chemistry*
  • Receptor, Adenosine A2A / metabolism
  • Receptors, G-Protein-Coupled / chemistry*
  • Receptors, G-Protein-Coupled / metabolism

Substances

  • Ligands
  • Receptor, Adenosine A2A
  • Receptors, G-Protein-Coupled