The overall objectives are to develop nuclear magnetic resonance (NMR) techniques and use them in concert with other experimental approaches to elucidate the molecular structure and physiologic functions of selected membrane-targeting proteins involved in phototransduction in vision and other signal transduction processes. During the next five years, we will use nuclear magnetic resonance (NMR), fluorescence, microcalorimetry, spin-label EPR, x-ray crystallography, and computational analysis to delineate the structure, dynamics and mechanisms of a family of neuronal calcium sensor proteins (calcium-myristoyl switches) that serve as membrane- targeting regulators in calcium signaling and are linked to retinal and neurological diseases. Our studies will focus on retinal recoverin, involved in cancer-associated retinopathy;guanylate cyclase activating proteins (GCAPs);linked to autosomal dominant cone dystrophy;centrins, that control protein translocation in photoreceptors;and calcium binding protein-4 (CaBP4), implicated in congenital stationary night blindness. By continuing our intensive study of retinal recoverin and the GCAP proteins and by broadening its scope to encompass neuronal homologs and protein targets, we hope to gain an atomic-level understanding of how calcium sensor proteins operate in signal transduction and disease processes. In particular, we want to understand how covalently attached myristoyl groups work in concert with calcium-binding sites and target proteins to guide this family of proteins to specific membrane-bound targets.
The specific aims are 3-fold: (1) Determine atomic-level structures of the GCAP proteins bound to retinal guanylate cyclases (RetGCs) to elucidate the Ca2+-dependent activation mechanism of RetGCs and thus provide a structural basis for understanding mechanisms of visual recovery and retinal degenerative diseases;(2) Determine structures of centrin-1 bound to T?? to provide a structural basis for understanding the mechanism of light-dependent protein translocation and its role in light adaptation;(3) Determine atomic-level structures of the retinal calcium sensor protein (CaBP4) and its structural interaction with voltage-gated Ca2+ channels (CaV1.4) at the rod synapse to understand the Ca2+-dependent regulatory mechanism of ion channels linked to retinal diseases.

Public Health Relevance

Calcium ion (Ca2+) in the cell is important for transmitting and regulating neural signals in vision. The goal of our research is to understand how calcium sensor proteins in the brain and retina transduce calcium signals and regulate the transport of cellular Ca2+ through ion channels during phototransduction and cell signaling.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY012347-16
Application #
8442262
Study Section
Special Emphasis Panel (ZRG1-BDPE-N (09))
Program Officer
Neuhold, Lisa
Project Start
1999-01-01
Project End
2016-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
16
Fiscal Year
2013
Total Cost
$321,160
Indirect Cost
$107,410
Name
University of California Davis
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
047120084
City
Davis
State
CA
Country
United States
Zip Code
95618
Ames, James B (2018) Dimerization of Neuronal Calcium Sensor Proteins. Front Mol Neurosci 11:397
Lim, Sunghyuk; Cudia, Diana; Yu, Qinhong et al. (2018) Chemical shift assignments of retinal degeneration 3 protein (RD3). Biomol NMR Assign 12:167-170
Lim, Sunghyuk; Roseman, Graham; Peshenko, Igor et al. (2018) Retinal guanylyl cyclase activating protein 1 forms a functional dimer. PLoS One 13:e0193947
Chowdhury, Dhrubajyoti; Turner, Matthew; Patriarchi, Tommaso et al. (2018) Ca2+/calmodulin binding to PSD-95 mediates homeostatic synaptic scaling down. EMBO J 37:122-138
Lim, Sunghyuk; Yu, Qinhong; Gottlieb, Sean M et al. (2018) Correlating structural and photochemical heterogeneity in cyanobacteriochrome NpR6012g4. Proc Natl Acad Sci U S A 115:4387-4392
Matt, Lucas; Kim, Karam; Hergarden, Anne C et al. (2018) ?-Actinin Anchors PSD-95 at Postsynaptic Sites. Neuron 97:1094-1109.e9
Li, Zhigang; Zhang, Yonghong; Hedman, Andrew C et al. (2017) Calmodulin Lobes Facilitate Dimerization and Activation of Estrogen Receptor-?. J Biol Chem 292:4614-4622
Lim, Sunghyuk; Scholten, Alexander; Manchala, Grace et al. (2017) Structural Characterization of Ferrous Ion Binding to Retinal Guanylate Cyclase Activator Protein 5 from Zebrafish Photoreceptors. Biochemistry 56:6652-6661
Yu, Qinhong; Lim, Sunghyuk; Rockwell, Nathan C et al. (2016) 1H, 15N, and 13C chemical shift assignments of cyanobacteriochrome NpR6012g4 in the red-absorbing dark state. Biomol NMR Assign 10:139-42
Li, Congmin; Lim, Sunghyuk; Braunewell, Karl H et al. (2016) Structure and Calcium Binding Properties of a Neuronal Calcium-Myristoyl Switch Protein, Visinin-Like Protein 3. PLoS One 11:e0165921

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