This is a Shared Instrumentation Grant proposal for the acquisition of a modern commercial CW and FT-EPR Bruker EleXsys X-band spectrometer for use by a core group of six established Principal Investigators and one beginning investigator from Universities in the Research Triangle area of North Carolina. Currently, there is no such instrument in the Triangle area. The ongoing research projects involve applications of modern time-domain EPR methods including pulsed saturation recovery and HYSCORE to a wide range of biomedical problems and systems from hybrid nanoscale assemblies for membrane protein biochips to lipid transfer proteins, self-assembly of tRNA and drug-delivery assisted by nanoparticle-templated virions, and the structures of the enzymes'catalytic sites. Currently, all these projects are either pursued with available continuous wave X-band EPR instruments that are at least 20 years old or in collaborating with other groups that have Bruker EleXsys X-band spectrometer available. With this proposed major upgrade all seven core research projects will benefit from recent developments in instrumentation and methodology of time-domain EPR methods at X-band. Specifically, we will be able to 1) obtain long-range (to up to 60-70 angstrom) distance constrains for large biomolecular assemblies and partially folded structures;2) directly measure electronic relaxation rates of specifically labeled proteins, RNAs, and phospholipids in the presence and absence of paramagnetic relaxation agents in order to gain valuable structural information;and 3) obtain unique information on paramagnetic metal ion coordination upon substrate binding to catalytic enzymes (HYSCORE experiment).

Public Health Relevance

The relevance of this project to public health will be in providing primarily NIH-funded investigators as well as other investigators from NCSU, UNC, and the Research Triangle area with an advanced FT- EPR spectrometer as these investigators develop new projects involving structure-function studies of proteins and RNAs. The new FT-EPR spectrometer will also enhance training of graduate students from the newly established NCSU RNA Biology/Chemistry -Biology Interface Training Program that involves ten NCSU Departments.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biomedical Research Support Shared Instrumentation Grants (S10)
Project #
1S10RR023614-01A2
Application #
7596125
Study Section
Special Emphasis Panel (ZRG1-BCMB-M (30))
Program Officer
Tingle, Marjorie
Project Start
2009-04-01
Project End
2011-03-31
Budget Start
2009-04-01
Budget End
2011-03-31
Support Year
1
Fiscal Year
2009
Total Cost
$500,000
Indirect Cost
Name
North Carolina State University Raleigh
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
042092122
City
Raleigh
State
NC
Country
United States
Zip Code
27695
McCombs, Nikolette L; Smirnova, Tatyana; Ghiladi, Reza A (2017) Oxidation of Pyrrole by Dehaloperoxidase-Hemoglobin: Chemoenzymatic Synthesis of Pyrrolin-2-Ones. Catal Sci Technol 7:3104-3118
Roberts, James G; Voinov, Maxim A; Schmidt, Andreas C et al. (2016) The Hydroxyl Radical is a Critical Intermediate in the Voltammetric Detection of Hydrogen Peroxide. J Am Chem Soc 138:2516-9
Smirnova, Tatyana I; Smirnov, Alex I (2015) Peptide-Membrane Interactions by Spin-Labeling EPR. Methods Enzymol 564:219-58
Voinov, Maxim A; Smirnov, Alex I (2015) Ionizable Nitroxides for Studying Local Electrostatic Properties of Lipid Bilayers and Protein Systems by EPR. Methods Enzymol 564:191-217
Barrios, David A; D'Antonio, Jennifer; McCombs, Nikolette L et al. (2014) Peroxygenase and oxidase activities of dehaloperoxidase-hemoglobin from Amphitrite ornata. J Am Chem Soc 136:7914-25
Dumarieh, Rania; D'Antonio, Jennifer; Deliz-Liang, Alexandria et al. (2013) Tyrosyl radicals in dehaloperoxidase: how nature deals with evolving an oxygen-binding globin to a biologically relevant peroxidase. J Biol Chem 288:33470-82
Voinov, Maxim A; Rivera-Rivera, Izarys; Smirnov, Alex I (2013) Surface electrostatics of lipid bilayers by EPR of a pH-sensitive spin-labeled lipid. Biophys J 104:106-16
Li, Jikun; Wang, Ke; Smirnova, Tatyana I et al. (2013) Isoprenoid biosynthesis: ferraoxetane or allyl anion mechanism for IspH catalysis? Angew Chem Int Ed Engl 52:6522-5
Zhao, Qingbiao; Nellutla, Saritha; Son, Won-Joon et al. (2011) Ba4KFe3O9: a novel ferrite containing discrete 6-membered rings of corner-sharing FeO4 tetrahedra. Inorg Chem 50:10310-8
Wang, Weixue; Wang, Ke; Li, Jikun et al. (2011) An ENDOR and HYSCORE investigation of a reaction intermediate in IspG (GcpE) catalysis. J Am Chem Soc 133:8400-3

Showing the most recent 10 out of 12 publications