Effects of phosphorylation on the structure of the G-protein receptor rhodopsin.

Upon activation by light, rhodopsin is subject to phosphorylation by rhodopsin kinase at serine and threonine residues in the carboxyl terminal region of the protein. A 19 amino acid peptide that corresponds to the carboxyl terminal end of rhodopsin (residues 330-348) and contains these phosphorylation sites was synthesized. The structure of this peptide was determined using two-dimensional proton NMR. The structure of this peptide was similar to that determined for this region in peptides corresponding to the carboxyl 33 and 43 amino acids of rhodopsin. The effect of phosphorylation on the structure of the carboxyl terminal domain of rhodopsin was determined by solving the solution structures of the peptide containing residues 330-348 with phosphorylation at one (residue 343), three (residues 343, 338, and 334) and seven residues (residues 334, 335, 336, 338, 340, 342, 343). These data indicate that the major structural change occurs upon phosphorylation of the first residue, and that an additional structural change occurs with seven phosphates.