10.1021/jm2003798.s001
Andrei Zhukov
Andrei
Zhukov
Stephen P. Andrews
Stephen P.
Andrews
James C. Errey
James C.
Errey
Nathan Robertson
Nathan
Robertson
Benjamin Tehan
Benjamin
Tehan
Jonathan S. Mason
Jonathan S.
Mason
Fiona H. Marshall
Fiona H.
Marshall
Malcolm Weir
Malcolm
Weir
Miles Congreve
Miles
Congreve
Biophysical Mapping of the Adenosine A<sub>2A</sub> Receptor
American Chemical Society
2015
BPM
SPR
surface plasmon resonance
binding site mutants
Biophysical Mapping
molecule binding site
2A
data
KD
ligand receptor interactions
2015-12-16 19:26:15
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Biophysical_Mapping_of_the_Adenosine_A_sub_2A_sub_Receptor/2016321
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 (<i>K</i><sub>D</sub>, <i>k</i><sub>on</sub>, and <i>k</i><sub>off</sub>) 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<sub>2A</sub> receptor are presented.