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      Site-directed mutagenesis of the human adenosine A 2A receptor. Critical involvement of Glu 13 in agonist recognition

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          Abstract

          A glutamic acid residue in the first transmembrane domain of the human adenosine A 2A receptor was mutated to glutamine. Radioligand binding studies on COS-7 cell membranes expressing either the wild-type or the mutant receptor revealed that the affinity of the prototypic agonist CGS21680 (2-[4-[(2-carboxyethyl)phenyl]ethylamino]-5′- N-ethylcarboxamidoadenosine) for the mutant receptor was 15-fold lower than for the wild-type receptor. This was confirmed in functional studies with intact cells. The EC 50 values of CGS21680 for the stimulation of cAMP production differed in a similar way. Antagonists of various chemical structure were equally effective on both mutant and wild-type receptors, thus the mutation selectively diminishes agonist affinity. We propose an indirect perturbation of the binding site, perhaps through a proton transfer mechanism as suggested by molecular modelling.

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          Author and article information

          Journal
          0101032
          1015
          Biochem Pharmacol
          Biochem. Pharmacol.
          Biochemical pharmacology
          0006-2952
          1873-2968
          4 July 2017
          01 September 2000
          23 August 2017
          : 60
          : 5
          : 661-668
          Affiliations
          [a ]Division of Medicinal Chemistry, Leiden/Amsterdam Center for Drug Research, PO Box 9502, 2300 RA Leiden, Netherlands
          [b ]Molecular Recognition Section, Laboratory of Bioorganic Chemistry, NIDDK, NIH, Bethesda, MD 20982, USA
          Author notes
          [* ]Corresponding author: LACDR/Medicinal Chemistry, PO Box 9502, 2300 RA Leiden, Netherlands. Tel.: +31 71 5274651; fax: +31 71 5274277
          Article
          PMC5567773 PMC5567773 5567773 nihpa890072
          5567773
          10927024
          3b165129-cee8-460f-96bf-54b8a0965ea7
          History
          Categories
          Article

          Adenosine A2A receptor,human,Mutation,Glutamic acid,CGS21680,XAC,Amiloride

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