The long-term objective of the proposed studies is to understand the biochemical steps in phototransduction, leading from rhodopsin activation, through its inactivation, and to its regeneration. The importance of inactivation and regeneration of photolyzed rhodopsin has become apparent recently, as malfunctions of these biochemical events during phototransduction lead to degeneration of photoreceptors and impairment of vision. Arrestin is an important component of inactivation of phototransduction and a potent auto-antigen. Elucidation of the structure and function of arrestin will greatly enhance our understanding of the pathogenesis of disease involving dysfunction of this protein. The applicant proposes to examine the structural properties of arrestin and P44 (a splice variant of arrestin) and how they participate in the quanching of phototransduction and in the visual cycle. The applicant seeks to understand: (1) the functional and structural domains of arrestin, continuing his crystallographic approaches; (2) the functional properties of truncated arrestin found in Oguchi Disease, and the molecular basis of arrestin heterogeneity; (3) the functional differences between arrestin and p44 using electrophysiological and biochemical methods; (4) which regions in the primary sequence of arrestin and p44 interact with photolyzed rhodopsin, phosphorylated at physiologically relevant sites (Ser338, Ser343, and Ser334); and finally (5) inactivation of photolyzed rhodopsin and steps in the visual cycle of mutant mice in which the rhodopsin kinase or arrestin genes were disrupted by targeted mutagenesis.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY009339-08
Application #
2634420
Study Section
Special Emphasis Panel (ZRG1-VISC (01))
Project Start
1992-08-01
Project End
2001-12-31
Budget Start
1998-01-01
Budget End
1998-12-31
Support Year
8
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Washington
Department
Ophthalmology
Type
Schools of Medicine
DUNS #
135646524
City
Seattle
State
WA
Country
United States
Zip Code
98195
Daruwalla, Anahita; Choi, Elliot H; Palczewski, Krzysztof et al. (2018) Structural biology of 11-cis-retinaldehyde production in the classical visual cycle. Biochem J 475:3171-3188
Gao, Songqi; Kahremany, Shirin; Zhang, Jianye et al. (2018) Retinal-chitosan Conjugates Effectively Deliver Active Chromophores to Retinal Photoreceptor Cells in Blind Mice and Dogs. Mol Pharmacol 93:438-452
Kiser, Philip D; Zhang, Jianye; Sharma, Aditya et al. (2018) Retinoid isomerase inhibitors impair but do not block mammalian cone photoreceptor function. J Gen Physiol 150:571-590
Luu, Jennings; Palczewski, Krzysztof (2018) Human aging and disease: Lessons from age-related macular degeneration. Proc Natl Acad Sci U S A 115:2866-2872
Gao, Songqi; Parmar, Tanu; Palczewska, Grazyna et al. (2018) Protective Effect of a Locked Retinal Chromophore Analog against Light-Induced Retinal Degeneration. Mol Pharmacol 94:1132-1144
Behnen, Petra; Felline, Angelo; Comitato, Antonella et al. (2018) A Small Chaperone Improves Folding and Routing of Rhodopsin Mutants Linked to Inherited Blindness. iScience 4:1-19
Chen, Yuanyuan; Chen, Yu; Jastrzebska, Beata et al. (2018) A novel small molecule chaperone of rod opsin and its potential therapy for retinal degeneration. Nat Commun 9:1976
Sui, Xuewu; Farquhar, Erik R; Hill, Hannah E et al. (2018) Preparation and characterization of metal-substituted carotenoid cleavage oxygenases. J Biol Inorg Chem 23:887-901
Gulati, Sahil; Jin, Hui; Masuho, Ikuo et al. (2018) Targeting G protein-coupled receptor signaling at the G protein level with a selective nanobody inhibitor. Nat Commun 9:1996
Kelly, Mary E; Ramkumar, Srinivasagan; Sun, Weizhong et al. (2018) The Biochemical Basis of Vitamin A Production from the Asymmetric Carotenoid ?-Cryptoxanthin. ACS Chem Biol 13:2121-2129

Showing the most recent 10 out of 211 publications