title: Precision vs flexibility in GPCR signaling.
abstract: The G protein coupled receptor (GPCR) rhodopsin activates the heterotrimeric G protein transducin (Gt) to transmit the light signal into retinal rod cells. The rhodopsin activity is virtually zero in the dark and jumps by more than one billion fold after photon capture. Such perfect switching implies both high fidelity and speed of rhodopsin/Gt coupling. We employed Fourier transform infrared (FTIR) spectroscopy and supporting all-atom molecular dynamics (MD) simulations to study the conformational diversity of rhodopsin in membrane environment and extend the static picture provided by the available crystal structures. The FTIR results show how the equilibria of inactive and active protein states of the receptor (so-called metarhodopsin states) are regulated by the highly conserved E(D)RY and Yx7K(R) motives. The MD data identify an intrinsically unstructured cytoplasmic loop region connecting transmembrane helices 5 and 6 (CL3) and show how each protein state is split into conformational substates. The C-termini of the Gtγ- and Gtα-subunits (GαCT and GγCT), prepared as synthetic peptides, are likely to bind sequentially and at different sites of the active receptor. The peptides have different effects on the receptor conformation. While GγCT stabilizes the active states but preserves CL3 flexibility, GαCT selectively stabilizes a single conformational substate with largely helical CL3, as it is found in crystal structures. Based on these results we propose a mechanism for the fast and precise signal transfer from rhodopsin to Gt, which assumes a stepwise and mutual reduction of their conformational space. The mechanism relies on conserved amino acids and may therefore underlie GPCR/G protein coupling in general.
authors.affiliation: Institut für Medizinische Physik und Biophysik (CC2), Charité-Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany. matthias.elgeti@charite.de
authors.forename: Matthias
authors.initials: M
authors.lastname: Elgeti
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Rose
authors.forename: Franz J
authors.initials: FJ
authors.lastname: Bartl
authors.forename: Peter W
authors.initials: PW
authors.lastname: Hildebrand
authors.forename: Klaus-Peter
authors.initials: KP
authors.lastname: Hofmann
authors.forename: Martin
authors.initials: M
authors.lastname: Heck
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011487:Protein Conformation; D012243:Rhodopsin / Q000819:agonists / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D017550:Spectroscopy, Fourier Transform Infrared; D015293:Transducin / Q000737:chemistry
chemical_list: D010446:Peptide Fragments; D012243:Rhodopsin; D015293:Transducin