IRBP enhances removal of 11- cis -retinaldehyde from isolated RPE membranes.
Interphotoreceptor retinoid-binding protein (IRBP) greatly enhances the conversion of all- trans -retinol to 11- cis -retinal by the retinal pigment epithelium (RPE) and facilitates 11- cis -retinal release from the RPE. However, the mechanisms by which IRBP exerts these effects are not clear. Using a model system of purified bovine IRBP and isolated bovine RPE membranes, we investigated the possibility that IRBP may favor the delivery of all- trans -retinol to, or the release of 11- cis -retinal from, RPE membranes. As the interphotoreceptor space contains serum retinol-binding protein (RBP) and serum albumin in addition to IRBP, we similarly examined the exchange of retinoids between RPE membranes and human RBP or bovine serum albumin (BSA). Isolated RPE membranes were loaded with radioactive 11- cis -retinal and incubated with solutions of IRBP, RBP, BSA or with buffer alone. Membranes (pellet) and retinoid-binding protein or buffer (supernatant) were separated by centrifugation and analysed for radioactive 11- cis -retinal. Membranes incubated with buffer alone released only 4-5% of their 11- cis -retinal, while 25 microm IRBP removed 18-35%. More retinal was released as the membrane concentration was reduced. In contrast, RBP and BSA removed little retinal, even though both proteins are capable of binding this retinoid. Similar results were obtained with bovine liver membranes, consistent with the idea that the effects of IRBP do not depend on an RPE surface receptor for IRBP. IRBP was also markedly superior to RBP and BSA in removing all- trans -retinol from RPE membranes. In addition, IRBP efficiently delivered bound all- trans -retinol to membranes; however, in contrast to their differential removal of retinoids, all three binding proteins delivered comparable amounts of retinol to membranes. (This result supports the practice of using BSA as a retinoid carrier in in vitro experimental systems). We conclude that, whereas IRBP shares with other retinoid-binding proteins the ability to deliver retinol to membranes, IRBP is unique in its capacity to remove 11- cis -retinal from membranes. This may be the feature of IRBP that drives the vitamin A cycle to efficiently produce 11- cis -retinal.
Edwards
R B
RB
Schepens Eye Research Institute, Boston, MA 02114, USA.
Adler
A J
AJ
D000818:Animals; D002417:Cattle; D005109:Extracellular Matrix; D005136:Eye Proteins; D006801:Humans; D010786:Photoreceptor Cells / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000378:metabolism*; D012172:Retinaldehyde / Q000378:metabolism*; D012177:Retinol-Binding Proteins / Q000494:pharmacology*; D000099214:Retinol-Binding Proteins, Interstitial
D005136:Eye Proteins; D012172:Retinaldehyde; D012177:Retinol-Binding Proteins; D000099214:Retinol-Binding Proteins, Interstitial