Abstract

In order properly to study many health-related problems it is necessary to understand how enzymes work. One of the best tools for doing this is kinetics, since in a kinetic study one looks at the reaction while it is taking place. Experience has shown that there are three levels of kinetic study of an enzyme. First, one establishes the kinetic mechanism, which is a description of the order in which reactants add to and products leave the enzyme. Second, one establishes the relative rates of the various steps in the kinetic mechanism. Finally, one attempts to deduce the actual chemical steps that occur on the enzyme. The purpose of this project is to develop kinetic tools for carrying out the various steps in analysis of the kinetic and chemical mechanism of enzymes, and to apply these tools to specific enzymes. Excellent progress towards these goals has been made, but all indications are that considerable development is still possible in the theoretical techniques, and of course, the number of enzymes whose kinetic and chemical mechanisms are well worked out is still small.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM018938-15
Application #
3269461
Study Section
Biochemistry Study Section (BIO)
Project Start
1977-01-01
Project End
1986-12-31
Budget Start
1986-01-01
Budget End
1986-12-31
Support Year
15
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
University of Wisconsin Madison
Department
Type
Earth Sciences/Resources
DUNS #
161202122
City
Madison
State
WI
Country
United States
Zip Code
53715

  Publications

Reinhardt, Laurie A; Thoden, James B; Peters, Greg S et al. (2013) pH-rate profiles support a general base mechanism for galactokinase (Lactococcus lactis). FEBS Lett 587:2876-81
Smith, Brian C; Anderson, Mark A; Hoadley, Kelly A et al. (2012) Structural and kinetic isotope effect studies of nicotinamidase (Pnc1) from Saccharomyces cerevisiae. Biochemistry 51:243-56
Thoden, James B; Reinhardt, Laurie A; Cook, Paul D et al. (2012) Catalytic mechanism of perosamine N-acetyltransferase revealed by high-resolution X-ray crystallographic studies and kinetic analyses. Biochemistry 51:3433-44
Saylor, Benjamin T; Reinhardt, Laurie A; Lu, Zhibing et al. (2012) A structural element that facilitates proton-coupled electron transfer in oxalate decarboxylase. Biochemistry 51:2911-20
Marlier, John F; Robins, Lori I; Tucker, Kathryn A et al. (2010) A kinetic and isotope effect investigation of the urease-catalyzed hydrolysis of hydroxyurea. Biochemistry 49:8213-9
Van Vleet, Jeremy; Kleeb, Andreas; Kast, Peter et al. (2010) 13C isotope effect on the reaction catalyzed by prephenate dehydratase. Biochim Biophys Acta 1804:752-4
Pinto-Tomas, Adrian A; Anderson, Mark A; Suen, Garret et al. (2009) Symbiotic nitrogen fixation in the fungus gardens of leaf-cutter ants. Science 326:1120-3
Marlier, John F; Fogle, Emily J; Cleland, W W (2008) A heavy-atom isotope effect and kinetic investigation of the hydrolysis of semicarbazide by urease from jack bean (Canavalia ensiformis). Biochemistry 47:11158-63
Van Vleet, Jeremy L; Reinhardt, Laurie A; Miller, Brian G et al. (2008) Carbon isotope effect study on orotidine 5'-monophosphate decarboxylase: support for an anionic intermediate. Biochemistry 47:798-803
Ralph, Erik C; Hirschi, Jennifer S; Anderson, Mark A et al. (2007) Insights into the mechanism of flavoprotein-catalyzed amine oxidation from nitrogen isotope effects on the reaction of N-methyltryptophan oxidase. Biochemistry 46:7655-64

Showing the most recent 10 out of 61 publications