Abstract

Phosphorylation of visual pigments is essential for normal vision. Deficiencies in this biochemical reaction can cause night blindness and photoreceptor apoptosis. Phosphorylation uncouples rhodopsin and most G-protein coupled receptors from downstream signaling pathways. The sites of phosphorylation on most G-protein coupled receptors are not known. During the previous funding period we developed a mass- spectrometry based method that enables us to determine the sites and kinetics of phosphorylation and dephosphorylation reactions that occur on visual pigments in intact retinas. We have used the method to establish the sites and kinetics of phosphorylation on mouse and zebrafish visual pigments under a variety of physiological conditions. ? ? We propose to use the method to pursue four new specific aims. ? ? 1. Evaluate rhodopsin phosphorylation during long-term illumination and identify relationships between visual pigment phosphorylation, visual cycle and visual sensitivity. ? ? 2. Evaluate factors that influence phosphorylation of cone visual pigments. ? ? 3. Identify structural characteristics that determine site selection and kinetics of phosphorylation by rhodopsin kinase. ? ? 4. Evaluate the influence of recoverin and Calcium on visual pigment phosphorylation. ? ? These studies will provide the most precise data yet available about the kinetics and site preferences for phosphorylation of G-protein coupled receptors. ? ? ?

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY006641-22
Application #
7279821
Study Section
Biology and Diseases of the Posterior Eye Study Section (BDPE)
Program Officer
Mariani, Andrew P
Project Start
1986-07-01
Project End
2011-06-30
Budget Start
2007-07-01
Budget End
2008-06-30
Support Year
22
Fiscal Year
2007
Total Cost
$374,394
Indirect Cost

  Institution

Name
University of Washington
Department
Biochemistry
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195

  Publications

Du, Jianhai; An, Jie; Linton, Jonathan D et al. (2018) How Excessive cGMP Impacts Metabolic Proteins in Retinas at the Onset of Degeneration. Adv Exp Med Biol 1074:289-295
Rajala, Ammaji; Wang, Yuhong; Brush, Richard S et al. (2018) Pyruvate kinase M2 regulates photoreceptor structure, function, and viability. Cell Death Dis 9:240
Zhu, Siyan; Yam, Michelle; Wang, Yekai et al. (2018) Impact of euthanasia, dissection and postmortem delay on metabolic profile in mouse retina and RPE/choroid. Exp Eye Res 174:113-120
Chao, Jennifer R; Knight, Kaitlen; Engel, Abbi L et al. (2017) Human retinal pigment epithelial cells prefer proline as a nutrient and transport metabolic intermediates to the retinal side. J Biol Chem 292:12895-12905
Kanow, Mark A; Giarmarco, Michelle M; Jankowski, Connor Sr et al. (2017) Biochemical adaptations of the retina and retinal pigment epithelium support a metabolic ecosystem in the vertebrate eye. Elife 6:
Hurley, James B (2017) Warburg's vision. Elife 6:
Du, Jianhai; Rountree, Austin; Cleghorn, Whitney M et al. (2016) Phototransduction Influences Metabolic Flux and Nucleotide Metabolism in Mouse Retina. J Biol Chem 291:4698-710
Du, Jianhai; Yanagida, Aya; Knight, Kaitlen et al. (2016) Reductive carboxylation is a major metabolic pathway in the retinal pigment epithelium. Proc Natl Acad Sci U S A 113:14710-14715
Zhang, Lijuan; Du, Jianhai; Justus, Sally et al. (2016) Reprogramming metabolism by targeting sirtuin 6 attenuates retinal degeneration. J Clin Invest 126:4659-4673
Contreras, Laura; Ramirez, Laura; Du, Jianhai et al. (2016) Deficient glucose and glutamine metabolism in Aralar/AGC1/Slc25a12 knockout mice contributes to altered visual function. Mol Vis 22:1198-1212

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