The broad aims of this work are to define the amplitudes and time scales of molecular motions that are crucial for biological function. These motions of interest include intramolecular side chain and backbone fluctuations as well as cooperative bending or breathing events. Translational motions are also crucial and include motions in pores or other confined spaces. Specifically, we propose to test in detail a new hypothesis that describes the control of proton spin-lattice relaxation in molecular solids including tissues that is important for interpretation of magnetic resonance images. We will use amide hydrogen exchange measurements in structurally well-characterized proteins to characterize the effects of stability on protein structural fluctuations. In particular we will measure the pressure and temperature dependence of amide hydrogen exchange rate constants to extract spatially resolved activation enthalpies, entropies, and volumes as a function of protein stability. These data will help define the character of structural fluctuations in proteins and define more clearly the character of the motions that permit reactions within the folded protein structure. We have demonstrated a simple experimental approach to mapping intermolecular contacts in solution using dioxygen and other paramagnetic co-solutes as the probe species. We will apply this experiment to map how oxygen contacts the surfaces of proteins and how it penetrates through folded protein structures. We will use a similar approach to examine solvation of small molecules and map experimentally the effects of electric charge on the nature of intermolecular contacts and compare the experimental maps with computational approaches to electric potential maps in folded proteins. We will apply NMR spin-lattice relaxation rate measurements in the rotating frame to measure protein intramolecular motions in the microsecond range and propose a similar experiment to define the water molecule lifetime distribution for long-lived water molecules that we may now count using magnetic relaxation dispersion measurements. Finally, we will extend work on molecular and nuclear spin dynamics of molecules in confined environments such as small tubes, pores, and locally ordered environments. This problem is important to understanding the spin relaxation in tissue systems, which is a valuable interpretative aid in clinical radiology.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM034541-17
Application #
6519168
Study Section
Biophysical Chemistry Study Section (BBCB)
Program Officer
Wehrle, Janna P
Project Start
1984-07-01
Project End
2005-03-31
Budget Start
2002-04-01
Budget End
2003-03-31
Support Year
17
Fiscal Year
2002
Total Cost
$254,214
Indirect Cost
Name
University of Virginia
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
001910777
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Teng, Ching-Ling; Hinderliter, Brian; Bryant, Robert G (2006) Oxygen accessibility to ribonuclease a: quantitative interpretation of nuclear spin relaxation induced by a freely diffusing paramagnet. J Phys Chem A 110:580-8
Teng, Ching-Ling; Bryant, Robert G (2004) Mapping oxygen accessibility to ribonuclease a using high-resolution NMR relaxation spectroscopy. Biophys J 86:1713-25
Teng, Ching-Ling; Martini, Silvia; Bryant, Robert G (2004) Local measures of intermolecular free energies in solution. J Am Chem Soc 126:15253-7
Victor, Ken G; Teng, Ching-Ling; Dinesen, T R D et al. (2004) Magnetic relaxation dispersion of lithium ion in solutions of DNA. Magn Reson Chem 42:518-23
Korb, Jean-Pierre; Bryant, Robert G (2002) Magnetic field dependence of proton spin-lattice relaxation times. Magn Reson Med 48:21-6
Teng, C L; Hong, H; Kiihne, S et al. (2001) Molecular oxygen spin-lattice relaxation in solutions measured by proton magnetic relaxation dispersion. J Magn Reson 148:31-4
Zhang, H; Lizitsa, N; Bryant, R G et al. (2001) Experimental characterization of intermolecular multiple-quantum coherence pumping efficiency in solution NMR. J Magn Reson 148:200-8
Dixon, M E; Hitchens, T K; Bryant, R G (2000) Comparisons of pressure and temperature activation parameters for amide hydrogen exchange in T4 lysozyme. Biochemistry 39:248-54
Kiihne, S; Bryant, R G (2000) Protein-bound water molecule counting by resolution of (1)H spin-lattice relaxation mechanisms. Biophys J 78:2163-9
Danek, A N; Bryant, R G (2000) Decay of dipolar order in diamagnetic and paramagnetic proteins and protein gels. J Magn Reson 143:35-8

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