Molecular Basis of Ligand Dissociation from the Adenosine A2A Receptor

Dong Guo; Albert C Pan; Ron O Dror; Tamara Mocking; Rongfang Liu; Laura H Heitman; David E Shaw; Adriaan P IJzerman

doi:10.1124/mol.115.102657

Molecular Basis of Ligand Dissociation from the Adenosine A2A Receptor

Mol Pharmacol. 2016 May;89(5):485-91. doi: 10.1124/mol.115.102657. Epub 2016 Feb 12.

Authors

Dong Guo¹, Albert C Pan¹, Ron O Dror¹, Tamara Mocking¹, Rongfang Liu¹, Laura H Heitman¹, David E Shaw¹, Adriaan P IJzerman²

Affiliations

¹ Division of Medicinal Chemistry, Leiden Academic Centre for Drug Research, Leidorken University, RA Leiden, The Netherlands (D.G., T.M., R.L., L.H.H., A.P.I.J.); D. E. Shaw Research, New York, New York (A.C.P., R.O.D., D.E.S.); and Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York (D.E.S.).
² Division of Medicinal Chemistry, Leiden Academic Centre for Drug Research, Leidorken University, RA Leiden, The Netherlands (D.G., T.M., R.L., L.H.H., A.P.I.J.); D. E. Shaw Research, New York, New York (A.C.P., R.O.D., D.E.S.); and Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York (D.E.S.) [email protected].

PMID: 26873858
DOI: 10.1124/mol.115.102657

Abstract

How drugs dissociate from their targets is largely unknown. We investigated the molecular basis of this process in the adenosine A2Areceptor (A2AR), a prototypical G protein-coupled receptor (GPCR). Through kinetic radioligand binding experiments, we characterized mutant receptors selected based on molecular dynamic simulations of the antagonist ZM241385 dissociating from the A2AR. We discovered mutations that dramatically altered the ligand's dissociation rate despite only marginally influencing its binding affinity, demonstrating that even receptor features with little contribution to affinity may prove critical to the dissociation process. Our results also suggest that ZM241385 follows a multistep dissociation pathway, consecutively interacting with distinct receptor regions, a mechanism that may also be common to many other GPCRs.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adenosine A2 Receptor Antagonists / chemistry
Adenosine A2 Receptor Antagonists / metabolism*
Adenosine A2 Receptor Antagonists / pharmacology
Amino Acid Substitution
Binding Sites
Cell Membrane / chemistry
Cell Membrane / drug effects
Cell Membrane / metabolism
Databases, Protein
HEK293 Cells
Humans
Hydrogen Bonding
Hydrophobic and Hydrophilic Interactions
Kinetics
Ligands
Models, Molecular*
Molecular Conformation
Molecular Dynamics Simulation
Mutation
Radioligand Assay
Receptor, Adenosine A2A / chemistry
Receptor, Adenosine A2A / genetics
Receptor, Adenosine A2A / metabolism*
Recombinant Proteins / chemistry
Recombinant Proteins / metabolism
Solubility
Triazines / chemistry
Triazines / metabolism*
Triazines / pharmacology
Triazoles / chemistry
Triazoles / metabolism*
Triazoles / pharmacology

Substances

Adenosine A2 Receptor Antagonists
Ligands
Receptor, Adenosine A2A
Recombinant Proteins
Triazines
Triazoles
ZM 241385