In the present study, a high resolution structure for the extramembranous regions of rhodopsin will be determined. The extramembranous domains will then be docked with the transmembrane helices of rhodopsin. At the present time, no high resolution structure is available for any G-protein receptor. Therefore, this study may provide the first such high resolution structure. These studies exploit the independent structural domains of the protein and represent an alternative method of determining integral membrane structure which may be generally applicable to membrane protein. Extensive literature on the independent stability of protein domains and the agreement between the structure determined here and measurements on intact rhodopsin supports this approach. Differential scanning calorimetry (DSC), circular dichroism (CD) and two-dimensional NMR will be used to determine the structures and stability of the protein domains. As rhodopsin is an integral membrane protein whose structure and function are influenced by the lipid bilayer, the role of both irreversibly-bound and reversibly-bound lipid on rhodopsin structure and stability will also be investigated. It is anticipated that this structure of rhodopsin will allow visualization of the binding site of transducin as well as the sites of phosphorylation. Furthermore, having determined the structure of the native rhodopsin, the structural consequences of mutations which are linked to autosomal dominant retinitis pigmentosa will be determined. These include mutations which are on both the extradiskal and intradiskal surfaces. Thus, rhodopsin structural defects could be linked to the disease state.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
2R01EY003328-15A1
Application #
2487832
Study Section
Visual Sciences C Study Section (VISC)
Project Start
1981-05-01
Project End
2001-03-31
Budget Start
1998-04-01
Budget End
1999-03-31
Support Year
15
Fiscal Year
1998
Total Cost
Indirect Cost
Name
University of Connecticut
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
City
Storrs-Mansfield
State
CT
Country
United States
Zip Code
06269
Yeagle, Philip L; Albert, Arlene D (2003) A conformational trigger for activation of a G protein by a G protein-coupled receptor. Biochemistry 42:1365-8
Boesze-Battaglia, Kathleen; Goldberg, Andrew F X; Dispoto, Janice et al. (2003) A soluble peripherin/Rds C-terminal polypeptide promotes membrane fusion and changes conformation upon membrane association. Exp Eye Res 77:505-14
Choi, Gregory; Landin, Judith; Galan, Jhenny Flor et al. (2002) Structural studies of metarhodopsin II, the activated form of the G-protein coupled receptor, rhodopsin. Biochemistry 41:7318-24
Katragadda, M; Chopra, A; Bennett, M et al. (2001) Structures of the transmembrane helices of the G-protein coupled receptor, rhodopsin. J Pept Res 58:79-89
Yeagle, P L; Choi, G; Albert, A D (2001) Studies on the structure of the G-protein-coupled receptor rhodopsin including the putative G-protein binding site in unactivated and activated forms. Biochemistry 40:11932-7
Katragadda, M; Alderfer, J L; Yeagle, P L (2001) Assembly of a polytopic membrane protein structure from the solution structures of overlapping peptide fragments of bacteriorhodopsin. Biophys J 81:1029-36
Landin, J S; Katragadda, M; Albert, A D (2001) Thermal destabilization of rhodopsin and opsin by proteolytic cleavage in bovine rod outer segment disk membranes. Biochemistry 40:11176-83
Young, J E; Albert, A D (2001) Rhodopsin palmitoylation in bovine rod outer segment disk membranes of different age/spatial location. Exp Eye Res 73:735-7
Yeagle, P L; Salloum, A; Chopra, A et al. (2000) Structures of the intradiskal loops and amino terminus of the G-protein receptor, rhodopsin. J Pept Res 55:455-65
Katragadda, M; Alderfer, J L; Yeagle, P L (2000) Solution structure of the loops of bacteriorhodopsin closely resembles the crystal structure. Biochim Biophys Acta 1466:6-Jan

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