Effect of light on endogenous ligands carried by interphotoreceptor retinoid-binding protein.
Interphotoreceptor retinoid-binding protein (IRBP) is a vitamin A carrier present only in the extracellular material lying between the neural retina and the retinal pigment epithelium of vertebrate eyes. The amount of retinol bound endogenously by IRBP in this interphotoreceptor space is known to increase upon illumination. This finding led to the hypothesis that IRBP may act as a shuttle for vitamin A during the visual cycle that regenerates rhodopsin. In the present work, we separated IRBP from other retinoid-binding proteins in bovine interphotoreceptor matrix preparations by means of size-exclusion chromatography. IRBP's endogenous ligands were retained during this procedure and were then extracted into hexane and analysed by normal-phase HPCL. We found that IRBP carries, in a light-dependent manner, all the retinoid isomers involved in the visual cycle. For dark-adapted eyes the amounts of bound ligands are (in nmol per eye) 0.09 all-trans retinol, 0.11 11-cis retinol, 0.04 all-trans retinal, 0.16 11-cis retinal, and 0.07 retinyl esters. For light-adapted eyes the amount of all-trans retinol was found to increase by a factor of five, and that of 11-cis retinal to decrease by a factor of four. (These eyes contain 3.1 nmol of IRBP, which does not change in amount with lighting conditions). Thus, the major endogenous ligand of IRBP is 11-cis retinaldehyde in the dark and all-trans retinol in the light. The data are consistent with a role for IRBP as a non-selective scavenger and stabilizer of retinoids released from photoreceptors and pigment epithelial cells. However, it cannot be concluded from these data that IRBP is involved in directed transport of retinoids across the interphotoreceptor matrix, since there is no evidence for the appropriate spatial gradients in the ligands bound to the protein.
Eye Research Institute, Boston, MA 02114.
A J
AJ
Adler
S A
SA
Spencer
D000221:Adaptation, Ocular / Q000502:physiology; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003623:Dark Adaptation / Q000502:physiology; D005136:Eye Proteins / Q000378:metabolism*; D066298:In Vitro Techniques; D008027:Light; D012160:Retina / Q000378:metabolism*; D012176:Retinoids / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D000099214:Retinol-Binding Proteins, Interstitial
D005136:Eye Proteins; D012176:Retinoids; D012177:Retinol-Binding Proteins; D000099214:Retinol-Binding Proteins, Interstitial