Schiff-base deprotonation is mandatory for light-dependent rhodopsin phosphorylation.
The absorption of light by rhodopsin leads to the formation of an activated intermediate (R*) capable of catalysing the exchange of GTP for GDP in a retinal guanine-nucleotide-binding regulatory protein (transducin). The ability of R* to function as a catalyst is terminated by the rhodopsin kinase. The 10 nonactive-site lysine residues of rhodopsin can be reductively dimethylated to form permethylated rhodopsin (PMRh). This derivative is phosphorylated to the same extent as rhodopsin after photolysis. The monomethylation of the active-site lysine residue of PMRh yields active-site-methylated rhodopsin (AMRh). It had previously been shown, by using AMRh, that the formation of R* and its spectroscopic signature metarhodopsin II requires the photochemically induced deprotonation of the active-site Schiff base [Longstaff, Calhoon & Rando (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 4209-4213]. Here it is demonstrated that active-site Schiff-base deprotonation is also mandatory in the formation of the form of photolyzed rhodopsin that is susceptible to phosphorylation by rhodopsin kinase. In terms of the spectroscopically defined rhodopsin intermediates, this means that only metarhodopsin II and possibly metarhodopsin III are the actual substrates for rhodopsin kinase.
Seckler
B
B
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
Rando
R R
RR
D000818:Animals; D001665:Binding Sites; D003624:Darkness; D005136:Eye Proteins; D051606:G-Protein-Coupled Receptor Kinase 1; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D007700:Kinetics; D008027:Light; D008623:Mercaptoethanol / Q000494:pharmacology; D008745:Methylation; D010766:Phosphorylation; D010786:Photoreceptor Cells / Q000201:enzymology*; D011494:Protein Kinases / Q000378:metabolism*; D012168:Retinal Pigments / Q000378:metabolism*; D012243:Rhodopsin / Q000378:metabolism* / Q000528:radiation effects; D012374:Rod Cell Outer Segment / Q000201:enzymology*; D012545:Schiff Bases; D015293:Transducin / Q000378:metabolism; D009764:o-Phthalaldehyde / Q000494:pharmacology
D005136:Eye Proteins; D006898:Hydroxylamines; D012168:Retinal Pigments; D012545:Schiff Bases; D019811:Hydroxylamine; D008623:Mercaptoethanol; D009764:o-Phthalaldehyde; D012243:Rhodopsin; D011494:Protein Kinases; D051606:G-Protein-Coupled Receptor Kinase 1; D015293:Transducin