Surfaces of rod photoreceptor disk membranes: light-activated enzymes.
The light-activated GTP-binding protein (GBP) in toad rod outer segments has been located on the cytoplasmic surface (CS) of rod disk membranes by correlating biochemical results with images of quick-frozen, freeze-fractured, and deep-etched rod outer segments. This has been accomplished by selectively removing and replacing the 8-12-nm particles that are found on the CS of disk membranes, exactly in parallel with the GBP. In contrast, the large particles are not correlated with another major disk enzyme, the light-activated cGMP phosphodiesterase. We have been unable to visualize this protein. The surface density of large particles, one particle per eleven rhodopsins in isolated rod outer segments and one particle per nine rhodopsins in intact retina, correlates well with previous biochemical estimates of GBP numbers based on enzyme activity. After the identification of the large particles, we tested the effects of light on the density of particles on the surface of disk membranes in intact retinas. Retinas quick-frozen at various intervals after a bright flash of light show a modest increase (approximately 30%) in particle density by 10 s after the flash but no increase before 1 s. The number of particles on the disk membrane returns to dark levels between 1 and 10 min after the flash. The 1-s latency in the change of particle binding would appear to rule out this process as a mechanism for initiating phototransduction in the rod.
D015106:3',5'-Cyclic-GMP Phosphodiesterases / Q000032:analysis*; D000818:Animals; D001798:Blood Proteins / Q000032:analysis*; D002024:Bufo marinus; D005613:Freeze Etching; D005614:Freeze Fracturing; D019204:GTP-Binding Proteins; D008027:Light; D008854:Microscopy, Electron; D010786:Photoreceptor Cells / Q000201:enzymology*; D011956:Receptors, Cell Surface / Q000032:analysis*; D012374:Rod Cell Outer Segment / Q000201:enzymology* / Q000648:ultrastructure
D001798:Blood Proteins; D011956:Receptors, Cell Surface; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D019204:GTP-Binding Proteins