The proposed research will apply methods from synthetic and physical inorganic chemistry to selected transition metal proteins and enzymes possessing unusual structural features, as evidenced by their spectroscopic and catalytic properties. The general objective of the research is to understand how the chemical environment of the metal ions leads to their distinctive spectroscopic and catalytic properties. This will be accomplished by fundamental physical and chemical studies of an unusual iron-containing hydrolytic enzyme, and by the preparation of synthetic models for the interaction of iron-sulfur clusters with other chromophores in certain complex iron-sulfur enzymes. The specific examples to be investigated in this research are two biologically significant iron-containing systems: the purple phosphatases, and iron-sulfur enzymes with flavin or heme prosthetic groups. The purple acid phosphatase from bovine spleen contains a novel spin-coupled binuclear unit at or near the active site. Spectroscopic and chemical methods will be employed to determine the nature of this unusual prosthetic group in the bovine and other similar enzymes, and the role of the prosthetic group in catalysis of a simple hydrolytic (rather than an oxidation-reduction) reaction. Iron-sulfur clusters containing covalently bound flavin or porphyrin groups as ligands will be prepared and characterized, in order to allow an examination of the factors affecting electron transfer and magnetic interactions between these redox active groups. Many enzymes, as well as components of the mitochondrial electron transport chain, are known to contain and to transfer electrons between similar units.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM032117-04
Application #
3280729
Study Section
Metallobiochemistry Study Section (BMT)
Project Start
1983-01-01
Project End
1986-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
4
Fiscal Year
1986
Total Cost
Indirect Cost
Name
University of Virginia
Department
Type
Schools of Arts and Sciences
DUNS #
001910777
City
Charlottesville
State
VA
Country
United States
Zip Code
22904
Orlando, J L; Zirino, T; Quirk, B J et al. (1993) Purification and properties of the native form of the purple acid phosphatase from bovine spleen. Biochemistry 32:8120-9
Averill, B A; Vincent, J B (1993) Electronic absorption spectroscopy of nonheme iron proteins. Methods Enzymol 226:33-51
Wang, Z; Ming, L J; Que Jr, L et al. (1992) 1H NMR and NOE studies of the purple acid phosphatases from porcine uterus and bovine spleen. Biochemistry 31:5263-8
Vincent, J B; Crowder, M W; Averill, B A (1992) Multiple binding sites for tetrahedral oxyanion inhibitors of bovine spleen purple acid phosphatase. Biochemistry 31:3033-7
Vincent, J B; Crowder, M W; Averill, B A (1992) Hydrolysis of phosphate monoesters: a biological problem with multiple chemical solutions. Trends Biochem Sci 17:105-10
Crowder, M W; Vincent, J B; Averill, B A (1992) Electron paramagnetic resonance studies on the high-salt form of bovine spleen purple acid phosphatase. Biochemistry 31:9603-8
Vincent, J B; Crowder, M W; Averill, B A (1991) Evidence for a phosphoryl-enzyme intermediate in phosphate ester hydrolysis by purple acid phosphatase from bovine spleen. J Biol Chem 266:17737-40
Vincent, J B; Crowder, M W; Averill, B A (1991) Spectroscopic and kinetics studies of a high-salt-stabilized form of the purple acid phosphatase from bovine spleen. Biochemistry 30:3025-34
Vincent, J B; Averill, B A (1990) An enzyme with a double identity: purple acid phosphatase and tartrate-resistant acid phosphatase. FASEB J 4:3009-14
Vincent, J B; Averill, B A (1990) Sequence homology between purple acid phosphatases and phosphoprotein phosphatases. Are phosphoprotein phosphatases metalloproteins containing oxide-bridged dinuclear metal centers? FEBS Lett 263:265-8

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