title: A conserved carboxylic acid group mediates light-dependent proton uptake and signaling by rhodopsin.
abstract: A carboxylic acid residue is conserved at the cytoplasmic border of the third transmembrane segment among nearly all G protein-coupled receptors. In the visual receptor rhodopsin, replacement of the conserved Glu134 by a neutral glutamine results in enhanced transducin activation. Here we show that a key event in forming the active state of rhodopsin is proton uptake by Glu134 in the metarhodopsin II (MII) photoproduct. Site-directed mutants E134D and E134Q were studied by flash photolysis, where formation rates of their photoproducts and rates of pH change could be monitored simultaneously. Both mutants showed normal MII formation rates. However, E134D displayed a slowed rate of proton uptake and E134Q displayed a loss of light-induced uptake of two protons from the aqueous phase. Thus, Glu134 mediates light-dependent proton uptake by MII. We propose that receptor activation requires a light-induced conformational change that allows protonation of Glu134 and subsequent protonation of a second group. The strong conservation of Glu134 in G protein-coupled receptors implies a general requirement for a proton acceptor group at this position to allow light- or ligand-dependent receptor activation.
authors.affiliation: Institut für Biophysik und Strahlenbiologie, Universität Freiburg, Federal Republic of Germany.
authors.forename: S
authors.initials: S
authors.lastname: Arnis
authors.forename: K
authors.initials: K
authors.lastname: Fahmy
authors.forename: K P
authors.initials: KP
authors.lastname: Hofmann
authors.forename: T P
authors.initials: TP
authors.lastname: Sakmar
mesh_terms: D000595:Amino Acid Sequence; D002264:Carboxylic Acids / Q000378:metabolism*; D002460:Cell Line; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008027:Light; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011522:Protons; D012243:Rhodopsin / Q000031:analogs & derivatives / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D002264:Carboxylic Acids; D011522:Protons; C013110:metarhodopsins; D012243:Rhodopsin