Evaluation of the electron density profile of the frog rod outer segment disc-membrane in vivo using x-ray diffraction.

The structure of the rod outer segment disc-membrane in vivo was studied by X-ray low-angle scattering. The experiments were made on frogs under narcosis. Diffraction patterns corresponding to a resolution of 1.5 nm could be obtained from the membrane stacks of the rod outer segment discs. For the analysis of the measured diffraction pattern a new special computer procedure was elaborated. Among other generalizations of the theory, it was taken into account that the electron densities in the inter- and intra-disc spaces differ from the mean electron density of the whole stack. The consideration of this possibility, together with an exact experimental measurement of the isotrope background scattering, led to a mathematically unique solution. The calculated electron density profile apparently is a distinct asymmetric bilayer. The electron density of the side of the membrane which is in contact with the disc lumen is higher than the electron density on the side in contact with the cytoplasm. Therefore, a localization of rhodopsin or of other high molecular proteins mainly on the cytoplasmatic edge of the membrane can be excluded for the rod outer segment discs in vivo.