The focus of this proposal is on the structure and function of the visual pigment rhodopsin. There are two Specific Aims: 1. The 3-dimensional crystallization and x-ray structure determination of recombinant wild-type and mutant rhodopsins. While the x-ray crystal structure of bovine rhodopsin was solved several years ago, the protein used in that study was isolated from native sources. No one has as yet crystallized rhodopsin from a recombinant source. This is an important goal because once achieved it will allow a high resolution structural analysis of rhodopsin mutants. Structural analysis of rhodopsin mutants is a crucial next step for a fundamental understanding of the functioning of this protein. The mutants of greatest interest in this work will be those found in the retinal diseases retinitis pigmentosa and congenital night blindness, specifically those that constitutively activate the protein. 2. To use transgenic Xenopus laevis to explore the molecular mechanisms of retinal disease arising from mutations in the genes for rhodopsin and its downstream signaling component transducin. The main focus of this Aim will be on the disease, retinitis pigmentosa.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY007965-20
Application #
7344674
Study Section
Special Emphasis Panel (ZRG1-VISC (01))
Program Officer
Mariani, Andrew P
Project Start
1988-12-01
Project End
2009-11-30
Budget Start
2007-12-01
Budget End
2009-11-30
Support Year
20
Fiscal Year
2008
Total Cost
$366,369
Indirect Cost
Name
Brandeis University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
616845814
City
Waltham
State
MA
Country
United States
Zip Code
02454
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Devine, Erin L; Oprian, Daniel D; Theobald, Douglas L (2013) Relocating the active-site lysine in rhodopsin and implications for evolution of retinylidene proteins. Proc Natl Acad Sci U S A 110:13351-5
Deupi, Xavier; Edwards, Patricia; Singhal, Ankita et al. (2012) Stabilized G protein binding site in the structure of constitutively active metarhodopsin-II. Proc Natl Acad Sci U S A 109:119-24
Standfuss, Jörg; Edwards, Patricia C; D'Antona, Aaron et al. (2011) The structural basis of agonist-induced activation in constitutively active rhodopsin. Nature 471:656-60

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