Modulation of GDP release from transducin by the conserved Glu134-Arg135 sequence in rhodopsin.
A superfamily of seven-transmembrane helix receptors catalyzes GDP release from heterotrimeric guanine nucleotide-binding proteins (G proteins) to initiate the intracellular signaling cascade. The photoreceptor rhodopsin is a prototypical member of the superfamily that activates the retinal G protein transducin (Gt). The cytoplasmic domain of rhodopsin binds and activates Gt, but residues that stimulate GDP release from Gt have not been identified until now. We show here that the abnormal signal transduction phenotypes of several different mutations affecting the highly conserved Glu134-Arg135 charge pair result from alteration of the GDP release step in the Gt activation cascade. We propose that Glu134 and Arg135 constitute the site that directly provides the signal from rhodopsin to activate GDP release from Gt. Because the Glu/Asp-Arg sequence occurs at a topologically identical location in most of the seven-transmembrane helix receptors, we propose that these residues constitute a switch for signal transfer.
Acharya
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Department of Molecular Cardiology, Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
Karnik
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D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002417:Cattle; D002522:Chlorocebus aethiops; D017124:Conserved Sequence; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006153:Guanosine Diphosphate / Q000378:metabolism*; D007700:Kinetics; D008961:Models, Structural; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D017354:Point Mutation; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012243:Rhodopsin / Q000737:chemistry* / Q000378:metabolism*; D015398:Signal Transduction; D013053:Spectrophotometry; D015293:Transducin / Q000378:metabolism*; D014162:Transfection
D010446:Peptide Fragments; D011994:Recombinant Proteins; D006153:Guanosine Diphosphate; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D012243:Rhodopsin; D015293:Transducin