Fourier Transform Infrared Study of the Rod Outer Segment Disk and Plasma Membranes of Vertebrate Retina

Phospholipid composition and structure of disk and plasma membranes purified from bovine rod outer segments (ROS) are examined using Fourier transform infrared spectroscopy. Vibrational data indicate that both disk and plasma membranes lack sphingophospholipids, in contrast to the lens membranes. The hydrocarbon chains of the disk lipids are unsaturated by a factor of 5 over the acyl chains of the plasma lipids. The plasma lipids with 3-fold higher cholesterol and 5-fold higher saturation melt at a higher temperature (26 degrees C) than the disk lipids which melt at 16 degrees C. The transition temperature decreases by more than 20 degrees C in going from disk lipids to disk membrane, indicating a large drop in the enthalpy of the ROS membrane-matrix, presumably due to enhanced rhodopsin-lipid interaction. The lipid composition predisposes the disk and plasma membranes to be fluid and structurally disordered (about 84%) around physiological temperature. The fluid phospholipid environment of the disk membrane (i.e., just a few degrees above subzero temperatures) is considered to be vital for the ROS photoreceptor function. The amide I band profile of rhodopsin indicates an extensive alpha-helical (53%) peptide chain, with little beta-sheet (21%) and beta-turns (18%) in ROS membranes. This structure and/or conformation is conserved between 0-60 degrees C even though disk and plasma lipids undergo a phase change. The H-D exchange data indicate that as much as 84% of the peptide residues of ROS membranes in partially bleached retinas is accessible to D2O solvent after 1 h.