The overall goals of the research supported by this grant are to develop NMR methods for determining the structures of proteins in biological supramolecular structures, and to apply NMR spectroscopy to the study of filamentous bacteriophage coat proteins in membranes and virus particles, including those that display peptides on their surface. Experimental methods and instrumentation will be developed so that highresolution NMR data can be obtained from all backbone and side chain sites of proteins in biological supramolecular structures. This will enable NMR spectroscopy to be used to determine the complete three-dimensional atomic resolution structures of proteins in virus particles, membranes, and other biological supramolecular structures. The structures of the major coat proteins of several different filamentous bacteriophages will be determined by solid-state NMR experiments on completely aligned bacteriophage particles, and the structures of the membrane-bound forms of these proteins will be determined in weakly aligned micelles by solution NMR and in completely aligned bilayers by solid-state NMR. Comparisons between the viral and membrane-bound forms of the protein will give insight into the assembly process. NMR will also be used to determine the structures of peptide sequences inserted near the N-terminus of the major pVIII coat protein and displayed on the surface of filamentous bacteriophage particles. The same filamentous bacteriophage coat protein that is used to generate the peptide sequences also provides an ideal platform for determining the structures of the peptides by the solid-state NMR methods that we are developing. The peptide sequences under investigation extend the biological of the research to AIDS, malaria, influenza, and neurochemistry.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB001966-29
Application #
7030988
Study Section
Biophysical Chemistry Study Section (BBCB)
Program Officer
Mclaughlin, Alan Charles
Project Start
1977-08-01
Project End
2008-02-28
Budget Start
2006-03-01
Budget End
2007-02-28
Support Year
29
Fiscal Year
2006
Total Cost
$315,434
Indirect Cost
Name
University of California San Diego
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
804355790
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Opella, Stanley J; Zeri, Ana Carolina; Park, Sang Ho (2008) Structure, dynamics, and assembly of filamentous bacteriophages by nuclear magnetic resonance spectroscopy. Annu Rev Phys Chem 59:635-57
Wu, Chin H; Opella, Stanley J (2008) Shiftless nuclear magnetic resonance spectroscopy. J Chem Phys 128:052312
Wu, Chin H; Opella, Stanley J (2008) Proton-detected separated local field spectroscopy. J Magn Reson 190:165-70
De Angelis, Anna A; Opella, Stanley J (2007) Bicelle samples for solid-state NMR of membrane proteins. Nat Protoc 2:2332-8
Grant, Christopher V; Sit, Siu-Ling; De Angelis, Anna A et al. (2007) An efficient (1)H/(31)P double-resonance solid-state NMR probe that utilizes a scroll coil. J Magn Reson 188:279-84
Sinha, Neeraj; Filipp, Fabian V; Jairam, Lena et al. (2007) Tailoring 13C labeling for triple-resonance solid-state NMR experiments on aligned samples of proteins. Magn Reson Chem 45 Suppl 1:S107-15
Sinha, Neeraj; Grant, Christopher V; Park, Sang Ho et al. (2007) Triple resonance experiments for aligned sample solid-state NMR of (13)C and (15)N labeled proteins. J Magn Reson 186:51-64
Thiriot, David S; Nevzorov, Alexander A; Opella, Stanley J (2005) Structural basis of the temperature transition of Pf1 bacteriophage. Protein Sci 14:1064-70
Thiriot, David S; Nevzorov, Alexander A; Zagyanskiy, Lena et al. (2004) Structure of the coat protein in Pf1 bacteriophage determined by solid-state NMR spectroscopy. J Mol Biol 341:869-79
Nevzorov, Alexander A; Mesleh, Michael F; Opella, Stanley J (2004) Structure determination of aligned samples of membrane proteins by NMR spectroscopy. Magn Reson Chem 42:162-71

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