Bovine Rhodopsin: Characterization of the Complex Formed with Triton X‐100

Abstract

The visual pigment rhodopsin from bovine retina has been purified devoid of lipids by use of the non‐ionic detergents Triton X‐100 and Emulphogen BC 720. Complete removal of lipids necessitates that rod outer segment membranes be dissociated by at least 10 times their weight of non‐ionic detergent. Rhodopsin can then be separated from other membrane proteins and lipids by ion‐exchange chromatography or density gradient sedimentation, provided that the detergent concentration is maintained at a level sufficient to ensure the dissociation of protein and lipid components. In the presence of micellar concentrations of detergent, Triton X‐100 binds to rhodopsin. Measurement of this binding and characterization of the complex show that it is composed of one rhodopsin and about 100 Triton X‐100 molecules. Photolysis of this complex results in a drastic reduction of the Triton X‐100 bound and an aggregation of the opsin formed. Copyright © 1974, Wiley Blackwell. All rights reserved