Funds for the purchase of a Bruker ElexSys E680 FT/CW spectrometer are requested. The spectrometer will seed the establishment of an advanced electron spin resonance (ESR) facility at Pitt, and will initially serve a group of nine investigators (Curran, Jen-Jacobson, Klein- Seetharaman, Saxena, Schafmeister, Trakselis, Waldeck, Wetzel, and Wipf). Seven of the users are supported by NIH grant types including P01, P50, R01, and R37 (MERIT). The primary appointments of the investigators are in Chemistry (Curran, Saxena, Trakselis, Waldeck, Wipf), Biology (Jen- Jacobson) and in the Medical School (Klein-Seetharaman, Wetzel) at the University of Pittsburgh, and in Chemistry (Schafmeister) at Temple University. Several of these investigators work collaboratively on the elucidation of structure-function relationships in membrane proteins and oligomeric proteins. The microscopic origins of macroscopic behavior of many of these systems cannot be easily understood using conventional structural tools, such as nuclear magnetic resonance or X-Ray diffraction. The spectrometer will be used to determine protein structure and dynamics, as well as electron transfer processes in chemical and biological materials. The high frequency spectrometer will: (a) allow sensitive measurement of fast dynamics in proteins and macromolecules;(b) enable high resolution measurement of nanometer range distances in biomolecules in order to determine constraints on macromolecular structure and conformational dynamics;(c) allow measurement of electron-nuclear distances in order to reveal metal-ion coordination in macromolecules;(d) monitor electron transfer and charge transfer processes in chemical and biological materials;and (e) enable enhanced resolution to the identity of transient radicals in order to elucidate organic reaction mechanisms. The user group requires more capacity for high-end pulsed ESR experiments, access to high frequency ESR experiments, as well as more sensitivity for concentration limited samples - these needs dictate the requested instrument. The Department of Chemistry will provide $175,000 as matching funds for the purchase of the spectrometer, as well as renovate and configure suitable space for the spectrometer in the Chevron Science Center (Chemistry Department). Biomedical Relevance: The spectrometer will support several projects that have strong biomedical relevance. These include the measurement of structure and conformational dynamics of several membrane proteins that are attractive drug targets, misfolding of several proteins implicated in neurodegenerative diseases, establishment of the general principles of protein-DNA interaction, as well as innovative synthetic chemistry directed towards drug discovery.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biomedical Research Support Shared Instrumentation Grants (S10)
Project #
1S10RR028701-01
Application #
7827737
Study Section
Special Emphasis Panel (ZRG1-IMST-B (32))
Program Officer
Levy, Abraham
Project Start
2011-05-01
Project End
2013-04-30
Budget Start
2011-05-01
Budget End
2013-04-30
Support Year
1
Fiscal Year
2011
Total Cost
$1,623,490
Indirect Cost
Name
University of Pittsburgh
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Cunningham, Timothy F; Pornsuwan, Soraya; Horne, W Seth et al. (2016) Rotameric preferences of a protein spin label at edge-strand β-sheet sites. Protein Sci 25:1049-60
De Santis, Emiliano; Minicozzi, Velia; Proux, Olivier et al. (2015) Cu(II)-Zn(II) Cross-Modulation in Amyloid-Beta Peptide Binding: An X-ray Absorption Spectroscopy Study. J Phys Chem B 119:15813-20
Kinde, Monica N; Chen, Qiang; Lawless, Matthew J et al. (2015) Conformational Changes Underlying Desensitization of the Pentameric Ligand-Gated Ion Channel ELIC. Structure 23:995-1004
Cunningham, Timothy F; Putterman, Miriam R; Desai, Astha et al. (2015) The double-histidine Cu²⁺-binding motif: a highly rigid, site-specific spin probe for electron spin resonance distance measurements. Angew Chem Int Ed Engl 54:6330-4
Cunningham, Timothy F; Shannon, Matthew D; Putterman, Miriam R et al. (2015) Cysteine-specific Cu2+ chelating tags used as paramagnetic probes in double electron electron resonance. J Phys Chem B 119:2839-43
Ji, Ming; Tan, Likun; Jen-Jacobson, Linda et al. (2014) Insights into copper coordination in the EcoRI-DNA complex by ESR spectroscopy. Mol Phys 112:3173-3182
Silva, K Ishara; Michael, Brian C; Geib, Steven J et al. (2014) ESEEM analysis of multi-histidine Cu(II)-coordination in model complexes, peptides, and amyloid-β. J Phys Chem B 118:8935-44
Silva, K Ishara; Saxena, Sunil (2013) Zn(II) ions substantially perturb Cu(II) ion coordination in amyloid-β at physiological pH. J Phys Chem B 117:9386-94