We wish to continue collaborative biological research demonstrating and utilizing the capabilities of our field-cycling device for use in shared commercial instruments. Our pneumatic device is relatively simple and is intended to be used in a variety of commercially available instruments, perhaps eventually at many other laboratories in some commercial form or as copies. Widely used commercial instruments are designed to work at some fixed field, where, for example, the sample's proton spins resonate at frequencies such as 500, or 600 MHz. Our new device allows researchers to program these experiments so that samples spend key parts of the time in lower fields, down to almost the earth's magnetic field. From this capability we get a better idea about time-scales and amplitudes of random-seeming motions in these molecules than cannot otherwise be obtained experimentally. In some cases these motions are key to their biological activity, while in others they may regulate these activities. Our results can then be used to help understand biochemical activity of enzymes, or to help validate computer simulations of their dynamics. Our device may also be useful for development of drugs, by helping to locate unknown activities in proteins, and their location in the structure. It has been applicable to a wide variety of molecules so far, including a short DMA duplex, an enzyme, and several phospholipids in membranes. Our plans include a major continuation of work (with Prof. Mary Roberts, Boston College) on membranes and how they interact with phospholipases; and several projects involving other collaborators, on several other enzymes. We will obtain new information about physical arrangements in these systems, and especially about dynamics of small molecules (substrates and inhibitors) weakly bound to enzymes.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM077974-04
Application #
7493388
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Chin, Jean
Project Start
2005-09-23
Project End
2010-08-31
Budget Start
2008-09-01
Budget End
2010-08-31
Support Year
4
Fiscal Year
2008
Total Cost
$146,020
Indirect Cost
Name
Brandeis University
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
616845814
City
Waltham
State
MA
Country
United States
Zip Code
02454
Redfield, Alfred G (2012) High-resolution NMR field-cycling device for full-range relaxation and structural studies of biopolymers on a shared commercial instrument. J Biomol NMR 52:159-77
Pu, Mingming; Orr, Andrew; Redfield, Alfred G et al. (2010) Defining specific lipid binding sites for a peripheral membrane protein in situ using subtesla field-cycling NMR. J Biol Chem 285:26916-22
Pu, Mingming; Fang, Xiaomin; Redfield, Alfred G et al. (2009) Correlation of vesicle binding and phospholipid dynamics with phospholipase C activity: insights into phosphatidylcholine activation and surface dilution inhibition. J Biol Chem 284:16099-107
Sivanandam, V N; Cai, Jingfei; Redfield, Alfred G et al. (2009) Phosphatidylcholine ""wobble"" in vesicles assessed by high-resolution 13C field cycling NMR spectroscopy. J Am Chem Soc 131:3420-1
Roberts, Mary F; Redfield, Alfred G; Mohanty, Udayan (2009) Phospholipid reorientation at the lipid/water interface measured by high resolution 31P field cycling NMR spectroscopy. Biophys J 97:132-41
Pu, Mingming; Feng, Jianwen; Redfield, Alfred G et al. (2009) Enzymology with a spin-labeled phospholipase C: soluble substrate binding by 31P NMR from 0.005 to 11.7 T. Biochemistry 48:8282-4
Clarkson, Michael W; Lei, Ming; Eisenmesser, Elan Z et al. (2009) Mesodynamics in the SARS nucleocapsid measured by NMR field cycling. J Biomol NMR 45:217-25
Wang, Yanling K; Chen, Wei; Blair, Derek et al. (2008) Insights into the structural specificity of the cytotoxicity of 3-deoxyphosphatidylinositols. J Am Chem Soc 130:7746-55
Klauda, Jeffery B; Roberts, Mary F; Redfield, Alfred G et al. (2008) Rotation of lipids in membranes: molecular dynamics simulation, 31P spin-lattice relaxation, and rigid-body dynamics. Biophys J 94:3074-83