An evaluation of purity criteria for bovine rod outer segment membranes.
The purity criteria of bovine rod outer segments (ROS) purified by different procedures were evaluated. Bovine ROS were purified by flotation and/or sedimentation in a continuous concentration gradient of sucrose. The purity of the different fractions was then evaluated according to four purity criteria: (i) the A280/delta A500 ratio, (ii) the moles of phospholipid per mole of rhodopsin, (iii) the fatty acid composition, and (iv) the interfacial properties of ROS membranes. All the purity criteria, except the A280/delta A500 ratio, were found to be adequate. From our results, the A280/delta A500 ratio cannot be used alone to characterize ROS purity. Furthermore, the phospholipid-to-rhodopsin ratio appears as the best purity criterion because of its reliability, its higher sensitivity, and its ease of achievement. It is noteworthy that mechanical treatment of the retinas dramatically affects the purification of ROS.
D000818:Animals; D002417:Cattle; D002458:Cell Fractionation; D002499:Centrifugation, Density Gradient; D005227:Fatty Acids / Q000032:analysis; D007425:Intracellular Membranes / Q000032:analysis; D010743:Phospholipids / Q000032:analysis; D010786:Photoreceptor Cells / Q000648:ultrastructure*; D012243:Rhodopsin / Q000032:analysis; D012374:Rod Cell Outer Segment / Q000648:ultrastructure*; D013053:Spectrophotometry; D013056:Spectrophotometry, Ultraviolet; D013500:Surface Tension
D005227:Fatty Acids; D010743:Phospholipids; D012243:Rhodopsin