Abstract

Aim 3. Adapt the available high throughput biophysical characterization techniques for IMPs, evaluate the quality of IMP production and sample preparation, and correlate data. a. Electron Microscopy - develop protein/lipid/detergent combinations by automated EM for sample homogeneity and protein quality by single particle picking using a low voltage EM (LVEM). b. Nuclear Magnetic Resonance Spectroscopy- develop the use of NMR probes and 1D 1H, 2D 1H15N and site specific 2H and l5N labeling with TROSY NMR for characterizing and optimizing IMP preparations. c. X-ray - adapt current automated nanovolume crystallization and imaging technologies for IMP use and evaluate IMP quality for potential crystallization and diffraction characterization.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Specialized Center (P50)
Project #
5P50GM073197-04
Application #
7484102
Study Section
Special Emphasis Panel (ZGM1)
Project Start
Project End
Budget Start
2007-08-01
Budget End
2008-07-31
Support Year
4
Fiscal Year
2007
Total Cost
$252,110
Indirect Cost

  Institution

Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037

  Publications

Ishchenko, Andrii; Peng, Lingling; Zinovev, Egor et al. (2017) Chemically Stable Lipids for Membrane Protein Crystallization. Cryst Growth Des 17:3502-3511
Lamichhane, Rajan; Liu, Jeffrey J; Pauszek 3rd, Raymond F et al. (2017) Fluorophore Labeling, Nanodisc Reconstitution and Single-molecule Observation of a G Protein-coupled Receptor. Bio Protoc 7:
Eddy, Matthew T; Didenko, Tatiana; Stevens, Raymond C et al. (2016) ?2-Adrenergic Receptor Conformational Response to Fusion Protein in the Third Intracellular Loop. Structure 24:2190-2197
Rowe, Timothy B; Luo, Zhe-Xi; Ketcham, Richard A et al. (2016) X-ray computed tomography datasets for forensic analysis of vertebrate fossils. Sci Data 3:160040
Bennett, Brad C; Purdy, Michael D; Baker, Kent A et al. (2016) An electrostatic mechanism for Ca(2+)-mediated regulation of gap junction channels. Nat Commun 7:8770
Horst, Reto; Wüthrich, Kurt (2015) Micro-scale NMR Experiments for Monitoring the Optimization of Membrane Protein Solutions for Structural Biology. Bio Protoc 5:
O'Connor, Casey; White, Kate L; Doncescu, Nathalie et al. (2015) NMR structure and dynamics of the agonist dynorphin peptide bound to the human kappa opioid receptor. Proc Natl Acad Sci U S A 112:11852-7
Lamichhane, Rajan; Liu, Jeffrey J; Pljevaljcic, Goran et al. (2015) Single-molecule view of basal activity and activation mechanisms of the G protein-coupled receptor ?2AR. Proc Natl Acad Sci U S A 112:14254-9
Moeller, Arne; Lee, Sung Chang; Tao, Houchao et al. (2015) Distinct conformational spectrum of homologous multidrug ABC transporters. Structure 23:450-460
Fenalti, Gustavo; Abola, Enrique E; Wang, Chong et al. (2015) Fluorescence Recovery After Photobleaching in Lipidic Cubic Phase (LCP-FRAP): A Precrystallization Assay for Membrane Proteins. Methods Enzymol 557:417-37

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