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

Here we report an efficient method to generate multiple co-structures of the A 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...

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Bibliographic Details
Published inScientific reports Vol. 8; no. 1; p. 41
Main Authors Rucktooa, Prakash, Cheng, Robert K Y, Segala, Elena, Geng, Tian, Errey, James C, Brown, Giles A, Cooke, Robert M, Marshall, Fiona H, Doré, Andrew S
Format Journal Article
LanguageEnglish
Published England 08.01.2018
Subjects
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
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Summary:Here we report an efficient method to generate multiple co-structures of the A 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).
ISSN:2045-2322