Hydrophobic amino acids at the cytoplasmic ends of helices 3 and 6 of rhodopsin conjointly modulate transducin activation

Laia Bosch-Presegué, Laura Iarriccio, Mnica Aguil, Darwin Toledo, Eva Ramon, Arnau Cordomí, Pere Garriga

Research output: Contribution to journalArticleResearchpeer-review

5 Citations (Scopus)

Abstract

Rhodopsin is the visual photoreceptor responsible for dim light vision. This receptor is located in the rod cell of the retina and is a prototypical member of the G-protein-coupled receptor superfamily. The structural details underlying the molecular recognition event in transducin activation by photoactivated rhodopsin are of key interest to unravel the molecular mechanism of signal transduction in the retina. We constructed and expressed rhodopsin mutants in the second and third cytoplasmic domains of rhodopsin - where the natural amino acids were substituted by the human M3 acetylcholine muscarinic receptor homologous residues - in order to determine their potential involvement in G-protein recognition. These mutants showed normal chromophore formation and a similar photobleaching behavior than WT rhodopsin, but decreased thermal stability in the dark state. The single mutant V1383.53 and the multiple mutant containing V2275.62 and a combination of mutations at the cytoplasmic end of transmembrane helix 6 caused a reduction in transducin activation upon rhodopsin photoactivation. Furthermore, combination of mutants at the second and third cytoplasmic domains revealed a cooperative role, and partially restored transducin activation. The results indicate that hydrophobic interactions by V1383.53, V2275.62, V2506.33, V2546.37 and I2556.38 are critical for receptor activation and/or efficient rhodopsin-transducin interaction. © 2010 Elsevier Ltd. All rights reserved.
Original languageEnglish
Pages (from-to)142-149
JournalArchives of Biochemistry and Biophysics
Volume506
Issue number2
DOIs
Publication statusPublished - 15 Feb 2011

Keywords

  • G-protein activation
  • G-protein-coupled receptor
  • Muscarinic receptor
  • Rhodopsin
  • Visual phototransduction

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