The Analytical Core facility will provide an essential I ink between the structural and biochemical studies proposed and application to diabetes. Additionally, this facility will provide a centralized resource. that will exploit existing technologies, previously established in the Taylor, Scott, Newton, and Olefsky laboratories and the Multi-Scale Imaging Core to expand the number and types of assays available for each laboratory to study signaling in diabetes. Therefore, the core will provide an essential point of synergy where al 1 laboratories will work together and ask critical questions about the molecular mechanisms underlying the diabetic state. This core, overseen by Drs. Taylor and King, will consist of four functional sub-units;protein expression, crystallography, proteomics/phosphoproteomics, and peptide arrays. The Analytical Core will provide the researchers in this Program Project Grant with 1)so luble, purified, recombinant proteins for structure-function studies;2) a centralized resource for protein crystallization, data colle cation, phase determination, model building, and structure refinement;3) ma ss spectrometry services . to map phosphorylation sites on proteins, identify proteins from crude and purified cellular fractionation experiments, define temporal protein -protein interactions, and quantitate protein expression;and 4) p eptide array technology to probe proteins of interest in molecular detail t o define interaction sites between proteins and generate 'super-binder'peptides? that can be used to manipulate signaling.

Public Health Relevance

The Analytical Core has traditionally provided a centralized point of synergy to the individual laboratories of this Program Project Grant and provided researchers access to specialized techniques that have accelerated the research and been inst rumental in the generation of no vel tools fo r the greater biological community. In the coming funding period, we have streamlined our services to provide better access of the laboratories to purified proteins, protein crystallization resources, a variety of mass spectrometry techniques, and peptide arrays.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Program Projects (P01)
Project #
5P01DK054441-15
Application #
8640920
Study Section
Special Emphasis Panel (ZDK1)
Project Start
Project End
Budget Start
2014-04-01
Budget End
2015-03-31
Support Year
15
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Smith, F Donelson; Esseltine, Jessica L; Nygren, Patrick J et al. (2017) Local protein kinase A action proceeds through intact holoenzymes. Science 356:1288-1293
Sastri, Mira; Darshi, Manjula; Mackey, Mason et al. (2017) Sub-mitochondrial localization of the genetic-tagged mitochondrial intermembrane space-bridging components Mic19, Mic60 and Sam50. J Cell Sci 130:3248-3260
Nystoriak, Matthew A; Nieves-CintrĂ³n, Madeline; Patriarchi, Tommaso et al. (2017) Ser1928 phosphorylation by PKA stimulates the L-type Ca2+ channel CaV1.2 and vasoconstriction during acute hyperglycemia and diabetes. Sci Signal 10:
Li, Lei; Li, Jing; Drum, Benjamin M et al. (2017) Loss of AKAP150 promotes pathological remodelling and heart failure propensity by disrupting calcium cycling and contractile reserve. Cardiovasc Res 113:147-159
Ilouz, Ronit; Lev-Ram, Varda; Bushong, Eric A et al. (2017) Isoform-specific subcellular localization and function of protein kinase A identified by mosaic imaging of mouse brain. Elife 6:
Inupakutika, Madhuri A; Sengupta, Soham; Nechushtai, Rachel et al. (2017) Phylogenetic analysis of eukaryotic NEET proteins uncovers a link between a key gene duplication event and the evolution of vertebrates. Sci Rep 7:42571
Nygren, Patrick J; Mehta, Sohum; Schweppe, Devin K et al. (2017) Intrinsic disorder within AKAP79 fine-tunes anchored phosphatase activity toward substrates and drug sensitivity. Elife 6:
Aggarwal-Howarth, Stacey; Scott, John D (2017) Pseudoscaffolds and anchoring proteins: the difference is in the details. Biochem Soc Trans 45:371-379
Turnham, Rigney E; Scott, John D (2016) Protein kinase A catalytic subunit isoform PRKACA; History, function and physiology. Gene 577:101-8
Scott, John D; Newton, Alexandra C (2016) Bacterial spore coat protein kinases: A new twist to an old story. Proc Natl Acad Sci U S A 113:6811-2

Showing the most recent 10 out of 211 publications