The first step in vision is the absorption of light by the chromophore of visual pigments. Vitamin A (retinol) is the precursor for visual pigment chromophore and also has important roles in retina development. Vitamin A is delivered from liver, the main storage site, to the eye by retinol binding protein (RBP), a specialized vitamin A carrier protein in the blood. RBP effectively solubilizes vitamin A, a hydrophobic molecule, and protects it from enzymatic and oxidative damage. In humans, loss of RBP function impairs dark adaptation, and causes progressive atrophy of the retinal pigment epithelium (RPE) in the eye at young ages. Under conditions of vitamin A limitation at which wild-type mice behave normally, RBP knockout mice have rapid vision loss after merely a week of vitamin A deficiency. The RPE is the main cell type in the eye for vitamin A absorption and storage for vision. Overwhelming evidence from many independent laboratories over the past 30 years suggests the existence of a specific RBP receptor that mediates vitamin A uptake in the RPE. Study of the molecular mechanism of vitamin A uptake into RPE has been greatly hampered by the lack of knowledge about the identity of the RBP receptor, which turned out to be extremely difficult to purify or clone. We have identified the RBP receptor using a novel biochemical strategy followed by mass spectrometry and have demonstrated, using a variety of techniques, that it has the expected properties of the RBP receptor. The RBP receptor is a multitransmembrane-domain protein of previously unknown function and is not homologous to any protein of known function. It binds to RBP with high affinity and has robust vitamin A activity from the vitamin A/RBP complex. Unlike most other small molecule delivery systems in eukaryotic cells that depend on extracellular carrier proteins, vitamin A uptake by the RBP receptor does not depend on endocytosis of the carrier protein RBP. We have immunolocalized the RBP receptor to the basolateral membrane of the RPE, a location consistent with its role in interacting with RBP in the choroidal blood for vitamin A uptake. In this proposal, we will elucidate the molecular mechanism of this novel membrane transport system by systematically studying the interaction of RBP with the RBP receptor, the mechanism of vitamin A uptake by the RBP receptor and the intracellular mechanism for the RBP receptor-mediated cellular uptake of vitamin A. Since vitamin A is essential for vision, the human eye is highly sensitive to vitamin A deficiency or to the loss of RBP function. Insufficient vitamin A uptake by the eye can lead to defective dark adaptation, RPE atrophy and even severe blindness. Understanding the molecular mechanism of vitamin A uptake by the eye will have a significant impact on efforts to preserve human vision.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY018144-05
Application #
8129515
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Neuhold, Lisa
Project Start
2007-09-30
Project End
2013-08-31
Budget Start
2011-09-01
Budget End
2013-08-31
Support Year
5
Fiscal Year
2011
Total Cost
$329,314
Indirect Cost
Name
University of California Los Angeles
Department
Physiology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Kawaguchi, Riki; Zhong, Ming; Kassai, Miki et al. (2015) Vitamin A Transport Mechanism of the Multitransmembrane Cell-Surface Receptor STRA6. Membranes (Basel) 5:425-53
Zhong, Ming; Sun, Hui (2015) A genetic clog in the vitamin A transport machinery. Cell 161:435-437
Kawaguchi, Riki; Zhong, Ming; Kassai, Miki et al. (2013) Differential and isomer-specific modulation of vitamin A transport and the catalytic activities of the RBP receptor by retinoids. J Membr Biol 246:647-660
Zhong, Ming; Kawaguchi, Riki; Ter-Stepanian, Mariam et al. (2013) Vitamin A transport and the transmembrane pore in the cell-surface receptor for plasma retinol binding protein. PLoS One 8:e73838
Kawaguchi, Riki; Zhong, Ming; Sun, Hui (2013) Real-time analyses of retinol transport by the membrane receptor of plasma retinol binding protein. J Vis Exp :e50169
Zhong, Ming; Kawaguchi, Riki; Kassai, Miki et al. (2012) Retina, retinol, retinal and the natural history of vitamin A as a light sensor. Nutrients 4:2069-96
Sun, Hui (2012) Membrane receptors and transporters involved in the function and transport of vitamin A and its derivatives. Biochim Biophys Acta 1821:99-112
Kawaguchi, Riki; Zhong, Ming; Kassai, Miki et al. (2012) STRA6-catalyzed vitamin A influx, efflux, and exchange. J Membr Biol 245:731-45
Casey, Jillian; Kawaguchi, Riki; Morrissey, Maria et al. (2011) First implication of STRA6 mutations in isolated anophthalmia, microphthalmia, and coloboma: a new dimension to the STRA6 phenotype. Hum Mutat 32:1417-26
Kawaguchi, Riki; Yu, Jiamei; Ter-Stepanian, Mariam et al. (2011) Receptor-mediated cellular uptake mechanism that couples to intracellular storage. ACS Chem Biol 6:1041-51

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