title: Biophysical mapping of the adenosine A2A receptor.
abstract: A new approach to generating information on ligand receptor interactions within the binding pocket of G protein-coupled receptors has been developed, called Biophysical Mapping (BPM). Starting from a stabilized receptor (StaR), minimally engineered for thermostability, additional single mutations are then added at positions that could be involved in small molecule interactions. The StaR and a panel of binding site mutants are captured onto Biacore chips to enable characterization of the binding of small molecule ligands using surface plasmon resonance (SPR) measurement. A matrix of binding data for a set of ligands versus each active site mutation is then generated, providing specific affinity and kinetic information (K(D), k(on), and k(off)) of receptor-ligand interactions. This data set, in combination with molecular modeling and docking, is used to map the small molecule binding site for each class of compounds. Taken together, the many constraints provided by these data identify key protein-ligand interactions and allow the shape of the site to be refined to produce a high quality three-dimensional picture of ligand binding, thereby facilitating structure based drug design. Results of biophysical mapping of the adenosine A(2A) receptor are presented.
authors.affiliation: Heptares Therapeutics Limited, BioPark, Welwyn Garden City, Hertfordshire, UK.
authors.forename: Andrei
authors.initials: A
authors.lastname: Zhukov
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Andrews
authors.forename: James C
authors.initials: JC
authors.lastname: Errey
authors.forename: Nathan
authors.initials: N
authors.lastname: Robertson
authors.forename: Benjamin
authors.initials: B
authors.lastname: Tehan
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Mason
authors.forename: Fiona H
authors.initials: FH
authors.lastname: Marshall
authors.forename: Malcolm
authors.initials: M
authors.lastname: Weir
authors.forename: Miles
authors.initials: M
authors.lastname: Congreve
mesh_terms: D058917:Adenosine A2 Receptor Antagonists / Q000737:chemistry*; D001665:Binding Sites / Q000235:genetics; D015195:Drug Design; D008024:Ligands; D008958:Models, Molecular; D009154:Mutation; D011743:Pyrimidines / Q000737:chemistry; D043705:Receptor, Adenosine A2A / Q000737:chemistry* / Q000235:genetics; D054852:Small Molecule Libraries; D014227:Triazines / Q000737:chemistry; D014230:Triazoles / Q000737:chemistry; D014970:Xanthines / Q000737:chemistry
chemical_list: D058917:Adenosine A2 Receptor Antagonists; D008024:Ligands; D011743:Pyrimidines; D043705:Receptor, Adenosine A2A; D054852:Small Molecule Libraries; D014227:Triazines; D014230:Triazoles; D014970:Xanthines; C097270:ZM 241385; C539997:2-(2-furanyl)-7-(2-(4-(4-(2-methoxyethoxy)phenyl)-1-piperazinyl)ethyl)-7H-pyrazolo(4,3-e)(1,2,4)triazolo(1,5-c)pyrimidine-5-amine