Reversible changes in circular dichroism spectra of cattle rhodopsin and isorhodopsin

When the disk membrane of rod outer segment is treated with detergents, the alpha-band CD of rhodopsin decreases and the gamma-band CD increases. This tendency of CD change is most prominent in the purified rhodopsin in cholic acid obtained by the ammonium sulfate fractionation of disk membranes, and the gamma-band CD is three times larger than the alpha-band CD. The beta-band CD of rhodopsin is only slightly influenced by detergents. The gamma-band of isorhodopsin shows two CD bands, one negative and one positive. Both in rhodopsin and isorhodopsin the gamma-band CD is lost by light irradiation. It is supposed that both chromophore retinal and aromatic amino acid residues of opsin are responsible for the gamma-band CD. When ammonium sulfate is added to the sonicated disk membranes suspended in cholic acid solution, the alpha-band CD of rhodopsin decreases to about a third and the gamma-band CD increases remarkably. The CD spectrum goes back to the original one on eliminating ammonium sulfate from the solution with dialysis. However, the purified rhodopsin recovers native CD spectrum on addition of lipids extracted from disk membranes. The retinal-opsin interaction that induces optical activity depends upon the property of a local environment formed by lipid and detergent.