| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1From the Department of Ophthalmology, University of Washington School of Medicine, Seattle, Washington; 2Cole Eye Institute and Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio; 3Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York; and the 4Department of Biochemistry, University of Washington, Seattle, Washington.
PURPOSE. To characterize mechanisms of apical localization of visual cycle components in retinal pigment epithelium (RPE) by the identification of cellular retinaldehyde-binding protein (CRALBP) interaction partners.
METHODS. An overlay assay was used to detect interactions of CRALBP with components of RPE microsomes. Interacting proteins were identified with two-dimensional (2D)-PAGE and liquid chromatography tandem mass spectrometry (LC MS/MS). Protein interactions were characterized by affinity chromatography, peptide competition, and expression of protein domains. Protein colocalization in mouse retina was examined using double-label immunocytochemistry and confocal microscopy.
RESULTS. CRALBP bound to a 54-kDa protein in RPE microsomes, which was identified as ERM (ezrin, radixin, moesin)-binding phosphoprotein 50 (EBP50), a PDZ domain protein, also known as sodium/hydrogen exchanger regulatory factory type 1 (NHERF-1). EBP50 and ezrin in solubilized microsomes bound to CRALBP-agarose but not to a control agarose column. CRALBP bound to both recombinant PDZ domains of EBP50 but not to the C-terminal ezrin-binding domain. In outer retina, EBP50 and ezrin were localized to RPE and Müller apical processes. CRALBP was distributed throughout both RPE and Müller cells, including their apical processes.
CONCLUSIONS. ERM proteins are multivalent linkers that connect plasma membrane proteins with the cortical actin cytoskeleton. EBP50 interacts with ERM family members through a C-terminal domain and binds targets such as CRALBP through its PDZ domains, thus contributing to an apical localization of target proteins. Our results provide a structural basis for apical localization of a retinoid-processing complex in RPE cells and offer insight into the cell biology of retinoid processing and trafficking in RPE. (247 words)
This article has been cited by other articles:
|
|
 |
|
  X. He, J. Lobsiger, and A. Stocker Bothnia dystrophy is caused by domino-like rearrangements in cellular retinaldehyde-binding protein mutant R234W PNAS, November 3, 2009; 106(44): 18545 - 18550. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. M. Farjo, G. Moiseyev, Y. Takahashi, R. K. Crouch, and J.-x. Ma The 11-cis-Retinol Dehydrogenase Activity of RDH10 and Its Interaction with Visual Cycle Proteins Invest. Ophthalmol. Vis. Sci., November 1, 2009; 50(11): 5089 - 5097. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Lange, Y. Yafai, A. Reichenbach, P. Wiedemann, and W. Eichler Regulation of Pigment Epithelium-Derived Factor Production and Release by Retinal Glial (Muller) Cells under Hypoxia Invest. Ophthalmol. Vis. Sci., November 1, 2008; 49(11): 5161 - 5167. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Collery, S. McLoughlin, V. Vendrell, J. Finnegan, J. W. Crabb, J. C. Saari, and B. N. Kennedy Duplication and Divergence of Zebrafish CRALBP Genes Uncovers Novel Role for RPE- and Muller-CRALBP in Cone Vision Invest. Ophthalmol. Vis. Sci., September 1, 2008; 49(9): 3812 - 3820. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. A. Deeg, D. Pompetzki, A. J. Raith, S. M. Hauck, B. Amann, S. Suppmann, T. W. F. Goebel, U. Olazabal, H. Gerhards, S. Reese, et al. Identification and Functional Validation of Novel Autoantigens in Equine Uveitis Mol. Cell. Proteomics, August 1, 2006; 5(8): 1462 - 1470. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Liden and U. Eriksson Understanding Retinol Metabolism: Structure and Function of Retinol Dehydrogenases J. Biol. Chem., May 12, 2006; 281(19): 13001 - 13004. [Full Text] [PDF]
|
|
|
|
|
|
K. Cortese, F. Giordano, E. M. Surace, C. Venturi, A. Ballabio, C. Tacchetti, and V. Marigo The Ocular Albinism Type 1 (OA1) Gene Controls Melanosome Maturation and Size Invest. Ophthalmol. Vis. Sci., December 1, 2005; 46(12): 4358 - 4364. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O. Strauss The Retinal Pigment Epithelium in Visual Function Physiol Rev, July 1, 2005; 85(3): 845 - 881. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F B Kalapesi, M T Coroneo, and M A Hill Human ganglion cells express the alpha-2 adrenergic receptor: relevance to neuroprotection Br J Ophthalmol, June 1, 2005; 89(6): 758 - 763. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Shenolikar, J. W. Voltz, R. Cunningham, and E. J. Weinman Regulation of Ion Transport by the NHERF Family of PDZ Proteins Physiology, December 1, 2004; 19(6): 362 - 369. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. L. Bonilha, S. K. Bhattacharya, K. A. West, J. Sun, J. W. Crabb, M. E. Rayborn, and J. G. Hollyfield Proteomic Characterization of Isolated Retinal Pigment Epithelium Microvilli Mol. Cell. Proteomics, November 1, 2004; 3(11): 1119 - 1127. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Wu, A. Hasan, T. Liu, D. C. Teller, and J. W. Crabb Identification of CRALBP Ligand Interactions by Photoaffinity Labeling, Hydrogen/Deuterium Exchange, and Structural Modeling J. Biol. Chem., June 25, 2004; 279(26): 27357 - 27364. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
