Lipid protein interactions couple protonation to conformation in a conserved cytosolic domain of G protein-coupled receptors.
The visual photoreceptor rhodopsin is a prototypical class I (rhodopsin-like) G protein-coupled receptor. Photoisomerization of the covalently bound ligand 11-cis-retinal leads to restructuring of the cytosolic face of rhodopsin. The ensuing protonation of Glu-134 in the class-conserved D(E)RY motif at the C-terminal end of transmembrane helix-3 promotes the formation of the G protein-activating state. Using transmembrane segments derived from helix-3 of bovine rhodopsin, we show that lipid protein interactions play a key role in this cytosolic "proton switch." Infrared and fluorescence spectroscopic pK(a) determinations reveal that the D(E)RY motif is an autonomous functional module coupling side chain neutralization to conformation and helix positioning as evidenced by side chain to lipid headgroup Foerster resonance energy transfer. The free enthalpies of helix stabilization and hydrophobic burial of the neutral carboxyl shift the side chain pK(a) into the range typical of Glu-134 in photoactivated rhodopsin. The lipid-mediated coupling mechanism is independent of interhelical contacts allowing its conservation without interference with the diversity of ligand-specific interactions in class I G protein-coupled receptors.
Madathil
Sineej
S
Division of Biophysics, Institute of Radiochemistry, Forschungszentrum Dresden-Rossendorf, PF 510119, D-01314 Dresden, Germany.
Fahmy
Karim
K
D020816:Amino Acid Motifs; D000818:Animals; D002417:Cattle; D003600:Cytosol / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008055:Lipids / Q000737:chemistry*; D008823:Micelles; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry; D011522:Protons; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry*; D012243:Rhodopsin / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods
D008024:Ligands; D008055:Lipids; D008823:Micelles; D011506:Proteins; D011522:Protons; D043562:Receptors, G-Protein-Coupled; D012243:Rhodopsin
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