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Ligand binding and subtype selectivity of the human A(2A) adenosine receptor: identification and characterization of essential amino acid residues

Jaakola, Veli-Pekka; Lane, J Robert; Lin, Judy Y; Katritch, Vsevolod; Ijzerman, Adriaan P; Stevens, Raymond C

Authors

Veli-Pekka Jaakola

ROB LANE ROB.LANE@NOTTINGHAM.AC.UK
Associate Professor

Judy Y Lin

Vsevolod Katritch

Adriaan P Ijzerman

Raymond C Stevens



Abstract

The crystal structure of the human A(2A) adenosine receptor bound to the A(2A) receptor-specific antagonist, ZM241385, was recently determined at 2.6-A resolution. Surprisingly, the antagonist binds in an extended conformation, perpendicular to the plane of the membrane, and indicates a number of interactions unidentified before in ZM241385 recognition. To further understand the selectivity of ZM241385 for the human A(2A) adenosine receptor, we examined the effect of mutating amino acid residues within the binding cavity likely to have key interactions and that have not been previously examined. Mutation of Phe-168 to Ala abolishes both agonist and antagonist binding as well as receptor activity, whereas mutation of this residue to Trp or Tyr had only moderate effects. The Met-177 --> Ala mutation impeded antagonist but not agonist binding. Finally, the Leu-249 --> Ala mutant showed neither agonist nor antagonist binding affinity. From our results and previously published mutagenesis data, we conclude that conserved residues Phe-168(5.29), Glu-169(5.30), Asn-253(6.55), and Leu-249(6.51) play a central role in coordinating the bicyclic core present in both agonists and antagonists. By combining the analysis of the mutagenesis data with a comparison of the sequences of different adenosine receptor subtypes from different species, we predict that the interactions that determine subtype selectivity reside in the more divergent "upper" region of the binding cavity while the "lower" part of the binding cavity is conserved across adenosine receptor subtypes.

Journal Article Type Article
Publication Date Apr 23, 2010
Journal Journal of Biological Chemistry
Print ISSN 0021-9258
Publisher American Society for Biochemistry and Molecular Biology
Peer Reviewed Peer Reviewed
Volume 285
Issue 17
Pages 13032-13044
APA6 Citation Jaakola, V., Lane, J. R., Lin, J. Y., Katritch, V., Ijzerman, A. P., & Stevens, R. C. (2010). Ligand binding and subtype selectivity of the human A(2A) adenosine receptor: identification and characterization of essential amino acid residues. Journal of Biological Chemistry, 285(17), 13032-13044. https://doi.org/10.1074/jbc.M109.096974
DOI https://doi.org/10.1074/jbc.M109.096974
Publisher URL https://www.jbc.org/content/285/17/13032
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