Regulation of retinal transducin by C-terminal peptides of rhodopsin.
Transducin is a multi-subunit guanine-nucleotide-binding protein that mediates signal coupling between rhodopsin and cyclic GMP phosphodiesterase in retinal rod outer segments. Whereas the T alpha subunit of transducin binds guanine nucleotides and is the activator of the phosphodiesterase, the T beta gamma subunit may function to link physically T alpha with photolysed rhodopsin. In order to determine the binding sites of rhodopsin to transducin, we have synthesized eight peptides (Rhod-1 etc.) that correspond to the C-terminal regions of rhodopsin and to several external and one internal loop region. These peptides were tested for their inhibition of restored GTPase activity of purified transducin reconstituted into depleted rod-outer-segment disc membranes. A marked inhibition of GTPase activity was observed when transducin was pre-incubated with peptides Rhod-1, Rhod-2 and Rhod-3. These peptides correspond to opsin amino acid residues 332-339, 324-331 and 317-321 respectively. Peptides corresponding to the three external loop regions or to the C-terminal residues 341-348 did not inhibit reconsituted GTPase activity. Likewise, Rhod-8, a peptide corresponding to an internal loop region of rhodopsin, did not inhibit GTPase activity. These findings support the concept that these specific regions of the C-terminus of rhodopsin serve as recognition sites for transducin.
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D020558:GTP Phosphohydrolases / Q000037:antagonists & inhibitors; D006165:Guanylyl Imidodiphosphate / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology*; D012160:Retina / Q000187:drug effects / Q000378:metabolism*; D012168:Retinal Pigments / Q000494:pharmacology*; D012243:Rhodopsin / Q000494:pharmacology*; D012374:Rod Cell Outer Segment / Q000378:metabolism; D015293:Transducin
D008565:Membrane Proteins; D010455:Peptides; D012168:Retinal Pigments; D006165:Guanylyl Imidodiphosphate; D012243:Rhodopsin; D020558:GTP Phosphohydrolases; D015293:Transducin