Biophysical Mapping of the Adenosine A2A Receptor
Open Access
- 10 June 2011
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of Medicinal Chemistry
- Vol. 54 (13), 4312-4323
- https://doi.org/10.1021/jm2003798
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 (KD, kon, and koff) 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 A2A receptor are presented.Keywords
This publication has 43 references indexed in Scilit:
- Ligand Binding and Subtype Selectivity of the Human A2A Adenosine ReceptorPublished by Elsevier BV ,2010
- GPCR 3D homology models for ligand screening: Lessons learned from blind predictions of adenosine A2a receptor complexProteins: Structure, Function, and Bioinformatics, 2009
- Thermostabilization of the Neurotensin Receptor NTS1Journal of Molecular Biology, 2009
- Evaluation of Homology Modeling of G-Protein-Coupled Receptors in Light of the A2A Adenosine Receptor Crystallographic StructureJournal of Medicinal Chemistry, 2009
- Co-evolving stability and conformational homogeneity of the human adenosine A 2a receptorProceedings of the National Academy of Sciences of the United States of America, 2008
- Structure of a β1-adrenergic G-protein-coupled receptorNature, 2008
- Conformational thermostabilization of the β1-adrenergic receptor in a detergent-resistant formProceedings of the National Academy of Sciences of the United States of America, 2008
- The Protein Data BankNucleic Acids Research, 2000
- CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choiceNucleic Acids Research, 1994
- Comparative Protein Modelling by Satisfaction of Spatial RestraintsJournal of Molecular Biology, 1993