Net electric charge on photopigment molecules and frog retinal receptor disk membrane structure

The photopigment molecules in frog retinal receptor disk membranes protude some 50-65% of their molecular diameter ( approximately 42 A) into the aqueous surface layer of the disk membrane, depending on whether the photopigment is bleached, while the remainder is embedded in the lipid core of the membrane. In order to determine whether the presumably polar groups covering this surface protruding into the aqueous phase possessed net electric charge, we collected X-ray diffraction data from the photopigment molecules in wet pellets of oriented disk membranes as a function of the pH and ionic strength of the sedimentation medium. The Fourier analysis applied to this data provided average nearest neighbor separations for the photopigment molecules for their planar arrangement in the disk membranes. Changes in the average separation of photopigment molecule nearest neighbors as a function of pH, ionic strength, and photopigment bleaching indicated that photopigment molecules possess negative net electric charge, that this net electric charge occurs in the aqueous surface layer of the disk membrane, and that this net charge is reduced on photopigment bleaching. This polar portion of the photopigment molecule may thereby determine the location of the photopigment molecules relative to the lipid core and other photopigment molecules in the disk membrane. In addition, the orientation (dichroism) of the photopigment relative to an axis normal to the plane of the disk membrane and the bleaching-dependent "sinking" of the photopigment molecule into the lipid core of the disk membrane may be accounted for.