Much of the proposed research represents a novel application of solid-state 67Zn NMR applied to proteins and their model systems. Many of the proposed experiments will be directed towards understanding the structural basis for 67Zn NMR parameters (quadrupole couplings, shielding tensors and their relative orientation). Low temperature 67Zn experiments will be performed as a means of obtaining the spectroscopic data. These data, in turn, will provide a critical test to the surrogate probe strategy of replacing Zn+2 with Cd+2 to investigate metal binding sites in metalloproteins, while providing invaluable structural information about this important class of proteins. Specific systems to be examined include the inhibitory Zn+2 site in carboxypetidase-A, characterization of intermediates in carbonic anhydrase and the unique chemistry afforded some Zn sites in DNA binding proteins. Further, the PI will prepare new model systems where the coordination environment about Zn and Cd can be carefully controlled, contrasted and investigated by single crystal and powder 67Zn and 113Cd NMR methods. Further, we will continue to develop the ab initio MO calculations of metal ion quadrupole and shielding tensors. These calculations have proven to be helpful in developing structural models for metal binding sites of biological complexes, e.g., the early stages of mineralization.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
8R01EB002050-24
Application #
6611018
Study Section
Metallobiochemistry Study Section (BMT)
Program Officer
Mclaughlin, Alan Charles
Project Start
1979-04-01
Project End
2004-03-31
Budget Start
2003-04-01
Budget End
2004-03-31
Support Year
24
Fiscal Year
2003
Total Cost
$656,250
Indirect Cost
Name
Battelle Pacific Northwest Laboratories
Department
Type
DUNS #
032987476
City
Richland
State
WA
Country
United States
Zip Code
99352
Harbison, Gerard S (2015) Polarization of core orbitals and computation of nuclear quadrupole coupling constants using Gaussian basis sets. J Magn Reson 257:24-31
Lipton, Andrew S; Heck, Robert W; de Jong, Wibe A et al. (2009) Low temperature 65Cu NMR spectroscopy of the Cu+ site in azurin. J Am Chem Soc 131:13992-9
Lipton, Andrew S; Heck, Robert W; Hernick, Marcy et al. (2008) Residue ionization in LpxC directly observed by 67Zn NMR spectroscopy. J Am Chem Soc 130:12671-9
Lipton, A S; Heck, R W; Primak, S et al. (2008) Characterization of Mg2+ binding to the DNA repair protein apurinic/apyrimidic endonuclease 1 via solid-state 25Mg NMR spectroscopy. J Am Chem Soc 130:9332-41
Lipton, Andrew S; Morlok, Melissa M; Parkin, Gerard et al. (2008) 67Zn solid-state NMR spectroscopy of ([Tp But,Me]Zn(OH2))[HOB(C6F5)3]. the importance of the anion [HOB(C6F5)3]-. Inorg Chem 47:5184-9
Lipton, Andrew S; Heck, Robert W; Staeheli, Greg R et al. (2008) A QM/MM approach to interpreting 67Zn solid-state NMR data in zinc proteins. J Am Chem Soc 130:6224-30
Lipton, Andrew S; Ellis, Paul D (2007) Modeling the metal center of Cys4 zinc proteins. J Am Chem Soc 129:9192-200
Bowers, Geoffrey M; Lipton, Andrew S; Mueller, Karl T (2006) High-field QCPMG NMR of strontium nuclei in natural minerals. Solid State Nucl Magn Reson 29:95-103
Lipton, Andrew S; Heck, Robert W; Ellis, Paul D (2004) Zinc solid-state NMR spectroscopy of human carbonic anhydrase: implications for the enzymatic mechanism. J Am Chem Soc 126:4735-9