Abstract
The molecular organization of the bovine rod outer segment (ROS) disk was investigated with respect to localization of the visual pigment rhodopsin in the membrane. The study made use of preparations of intact disks and ROS fragments. We used the membrane impermeable reagent sodium isethionyl acetimidate hydrochloride and membrane permeable reagent ethyl acetimidate hydrochloride to label membrane amino groups. We found that half the rhodopsin amino groups were accessible to the membrane impermeable reagent; therefore, rhodopsin must protrude into the extradiskal space. A complementary experiment using fluorescein isothiocyanate confirmed these results. Freeze-fracture electron microscopy showed that the disk membrane is asymmetric and that the sidedness of our disk preparation is uniform.
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