title: Molecular Basis of Ligand Dissociation from the Adenosine A2A Receptor.
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.
authors.affiliation: 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.).
authors.forename: Dong
authors.initials: D
authors.lastname: Guo
authors.affiliation: 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.).
authors.forename: Albert C
authors.initials: AC
authors.lastname: Pan
authors.affiliation: 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.).
authors.forename: Ron O
authors.initials: RO
authors.lastname: Dror
authors.affiliation: 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.).
authors.forename: Tamara
authors.initials: T
authors.lastname: Mocking
authors.affiliation: 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.).
authors.forename: Rongfang
authors.initials: R
authors.lastname: Liu
authors.affiliation: 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.).
authors.forename: Laura H
authors.initials: LH
authors.lastname: Heitman
authors.affiliation: 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.).
authors.forename: David E
authors.initials: DE
authors.lastname: Shaw
authors.affiliation: 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.) ijzerman@lacdr.leidenuniv.nl.
authors.forename: Adriaan P
authors.initials: AP
authors.lastname: IJzerman
mesh_terms: D058917:Adenosine A2 Receptor Antagonists / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D019943:Amino Acid Substitution; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D030562:Databases, Protein; D057809:HEK293 Cells; D006801:Humans; D006860:Hydrogen Bonding; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008024:Ligands; D008958:Models, Molecular; D008968:Molecular Conformation; D056004:Molecular Dynamics Simulation; D009154:Mutation; D011869:Radioligand Assay; D043705:Receptor, Adenosine A2A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012995:Solubility; D014227:Triazines / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D014230:Triazoles / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D058917:Adenosine A2 Receptor Antagonists; D008024:Ligands; D043705:Receptor, Adenosine A2A; D011994:Recombinant Proteins; D014227:Triazines; D014230:Triazoles; C097270:ZM 241385