This proposal represents a multinuclear, multifield, and multiphase investigation of the metal sites in metalloproteins. The primary nuclei to be examined in this research are 113Cd, 67Zn, and 95Mo. Much of the proposed research is a continuation of a successful research program in 113Cd NMR spectroscopy. The experiments include single crystal experiments on new model compounds for the active site o carboxypeptidase and continued work using Cd2+ as a prosthetic spin-spy in protoporphyrin dependent protein, i.e. hemoglobin and chlorophyll. However, several new experiments are proposed. These include dynamic angle spinning (DAS) experiments which will average the second order quadrupole broadening to zero. A primary objective of the experiments, cross-polarization in combination with DAS, is to make the observation of quadrupolar nuclides (67Zn and 95Mo) in metalloproteins a practical experiment in the solid state. Such experiments, if proven successful, would provide a much needed direct spectroscopic approach to study the structure and bonding in this important class of metalloproteins. Further, results from such research will provide independent collaboration of the results based on the surrogate probe strategy, i.e. 113Cd nmr of those proteins where the Zn2+ ion has been replaced by Cd2+. Further, it is demonstrated that ab initio MO calculations of metal ion shielding tensors are consistent with our simple """"""""shielding rules"""""""". These rules came about as a result of our single crystal experiments. Based upon these results, we will also utilize these calculations as a guide to interpret our protein data.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
7R01GM026295-14
Application #
3273799
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1979-04-01
Project End
1996-02-28
Budget Start
1993-08-01
Budget End
1994-02-28
Support Year
14
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Battelle Pacific Northwest Laboratories
Department
Type
DUNS #
032987476
City
Richland
State
WA
Country
United States
Zip Code
99352
Lipton, Andrew S; Bergquist, Catherine; Parkin, Gerard et al. (2003) Solid-State 67Zn NMR spectroscopic studies and ab initio molecular orbital calculations on a synthetic analogue of carbonic anhydrase. J Am Chem Soc 125:3768-72
Lipton, Andrew S; Wright, Terri A; Bowman, Michael K et al. (2002) Solid-state (67)zn NMR spectroscopy in bioinorganic chemistry. Spectra of four- and six-coordinate zinc pyrazolylborate complexes obtained by management of proton relaxation rates with a paramagnetic dopant. J Am Chem Soc 124:5850-60
Lipton, Andrew S; Smith, Mark D; Adams, Richard D et al. (2002) 67Zn solid-state and single-crystal NMR spectroscopy and X-ray crystal structure of zinc formate dihydrate. J Am Chem Soc 124:410-4
Lipton, A S; Sears, J A; Ellis, P D (2001) A general strategy for the NMR observation of half-integer quadrupolar nuclei in dilute environments. J Magn Reson 151:48-59
Lipton, A S; Buchko, G W; Sears, J A et al. (2001) 67Zn solid-state NMR spectroscopy of the minimal dna binding domain of human nucleotide excision repair protein XPA. J Am Chem Soc 123:992-3
McAteer, K; Lipton, A S; Kennedy, M A et al. (1996) A multiphase 113Cd NMR investigation of metalloporphyrin reorientation in cadmium-substituted myoglobin. Solid State Nucl Magn Reson 7:229-38
Koons, J M; Ellis, P D (1995) Applicability of factor analysis in solid state NMR. Anal Chem 67:4309-15
Ellis, P D (1989) 113Cd NMR of Cd2+-substituted carboxypeptidase. Support for a hexa-coordinate metal ion in the presence of inhibitors. J Biol Chem 264:3108-10
Ellis, P D; Marchetti, P S; Strang, P et al. (1988) Cadmium-substituted skeletal troponin C metal binding investigations and sequence assignment of the cadmium-113 resonances. J Biol Chem 263:10284-8
Bhattacharyya, L; Marchetti, P S; Ellis, P D et al. (1987) Nuclear magnetic resonance investigation of cadmium 113 substituted pea and lentil lectins. J Biol Chem 262:5616-21

Showing the most recent 10 out of 11 publications