Stubbs G W, Saari J C, Futterman S
J Biol Chem. 1979 Sep 10;254(17):8529-33.
11-cis-Retinal-binding protein from bovine retina has been purified to apparent homogeneity by gel filtration ion exchange, and hydroxylapatite chromatography. The molecular weight determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 33,000. A value of 36,000 was obtained by gel filtration. With 11-cis-retinal bound to the protein a bleachable spectral peak with maximum absorption at 425 nm was observed. When isolated without addition of exogenous retinal the purified binding protein displayed absorbance maxima at 340 and 425 nm, indicating the occurrence of two ligands in addition to the typical protein absorbance. The endogenous ligands responsible for these maxima were tentatively identified as 11-cis-retinol and 11-cis-retinal, respectively. The binding protein did not cross-react with antibodies prepared against bovine opsin, and its amino acid composition was distinct from that of opsin. The amount of retinal-binding protein extracted was approximately 1 nmol/retina or 1 mol of binding protein/20 mol of opsin. Its specificity and relative abundance suggest that the 11-cis-retinal-binding protein may play a major role in the visual cycle.
