Structure and ligand-binding site characteristics of the human P2Y11 nucleotide receptor deduced from computational modelling and mutational analysis - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Biochemical Journal Année : 2007

Structure and ligand-binding site characteristics of the human P2Y11 nucleotide receptor deduced from computational modelling and mutational analysis

Jacques Zylberg
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Denise Ecke
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Bilha Fischer
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Georg Reiser
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Résumé

The P2Y 11}-receptor (P2Y 11}-R) is a less explored drug target. We computed a hP2Y 11}-R homology model with two templates, bovine-rhodopsin (2.6Å resolution) and a hP2Y 1}:ATP complex model. The hP2Y 11}-R model was refined using molecular dynamics calculations and validated by virtual screening methods, with an enrichment factor of 5. Furthermore, mutational analyses of R106, E186, R268, R307 and A313 confirmed the adequacy of our hP2Y 11}-R model and the computed ligand recognition mode. The E186A and R268A mutants reduced the potency of ATP by one and three orders of magnitude, respectively. The R106A and R307A mutants were functionally inactive. We propose that residues R106, R268, R307 and E186 are involved in ionic interactions with the phosphate moiety of ATP. R307 is possibly also H-bonded to N 6} of ATP via the backbone carbonyl. Activity of ATP at the F109I mutant revealed that the proposed π-stacking of F109 with the adenine ring is a minor interaction. The mutation A313N, which is part of a hydrophobic pocket in the vicinity of the ATP-C2-position, partially explains the high activity of 2-MeS-ATP at P2Y 1}-R as compared to the negligible activity at the P2Y 11}-R. Inactivity of ATP at the Y261A mutant implies that Y261 acts as a molecular switch, as in other G-protein coupled receptors. Moreover, analysis of cAMP responses seen with the mutants showed that the efficacy of coupling of the P2Y 11}-R to G s} is more variable than coupling to G q}. Our model also indicates that S206 forms an H-bond with Pγ and M310 interacts with the adenine moiety.

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Dates et versions

hal-00478698 , version 1 (30-04-2010)

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Jacques Zylberg, Denise Ecke, Bilha Fischer, Georg Reiser. Structure and ligand-binding site characteristics of the human P2Y11 nucleotide receptor deduced from computational modelling and mutational analysis. Biochemical Journal, 2007, 405 (2), pp.277-286. ⟨10.1042/BJ20061728⟩. ⟨hal-00478698⟩

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