Abstract

The overall goal of this project is to construct a detailed picture of (i) the active site regions where the three partial reactions comprising the PPDK reaction take place, (ii) the mechanisms by which these three partial reactions take place and (iii) the mechanism by which the putative histidine phosphoryl carrier shuttles from one active site region to the next.
The specific aims of the project are: (1) To sequence the Bacteriodes symbiosus PPDK gene. (2) To crystallize PPDK and initiate X-ray crystallographic studies. (3) To determine the rate constants for the PPDK substrate binding and release steps and catalytic steps and to construct a free energy profile for the PPDK reaction. (4) To determine the stoichiometry, location and role of the metal ion cofactors during PPDK catalysis. (5) To identify the PPDK active site amino acid residues involved in substrate binding and catalysis. (6) To examine the structure of the active with regard to overlapping vs nonoverlapping sites for the three partial reactions and to test for movement of the putative """"""""phosphoryl carrier histidine"""""""" between reaction sites during catalysis.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM036260-07
Application #
3289857
Study Section
Biochemistry Study Section (BIO)
Project Start
1986-01-01
Project End
1995-08-31
Budget Start
1992-09-01
Budget End
1993-08-31
Support Year
7
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
University of Maryland College Park
Department
Type
Schools of Earth Sciences/Natur
DUNS #
City
College Park
State
MD
Country
United States
Zip Code
20742

  Publications

Wu, Chun; Dunaway-Mariano, Debra; Mariano, Patrick S (2013) Design, synthesis, and evaluation of inhibitors of pyruvate phosphate dikinase. J Org Chem 78:1910-22
Reger, Albert S; Wu, Rui; Dunaway-Mariano, Debra et al. (2008) Structural characterization of a 140 degrees domain movement in the two-step reaction catalyzed by 4-chlorobenzoate:CoA ligase. Biochemistry 47:8016-25
Wu, Rui; Cao, Jian; Lu, Xuefeng et al. (2008) Mechanism of 4-chlorobenzoate:coenzyme a ligase catalysis. Biochemistry 47:8026-39
Lim, Kap; Read, Randy J; Chen, Celia C H et al. (2007) Swiveling domain mechanism in pyruvate phosphate dikinase. Biochemistry 46:14845-53
Chen, Celia C H; Han, Ying; Niu, Weiling et al. (2006) Structure and kinetics of phosphonopyruvate hydrolase from Variovorax sp. Pal2: new insight into the divergence of catalysis within the PEP mutase/isocitrate lyase superfamily. Biochemistry 45:11491-504
Lin, Ying; Lusin, Jacqueline D; Ye, Dongmei et al. (2006) Examination of the structure, stability, and catalytic potential in the engineered phosphoryl carrier domain of pyruvate phosphate dikinase. Biochemistry 45:1702-11
Teplyakov, Alexey; Liu, Sijiu; Lu, Zhibing et al. (2005) Crystal structure of the petal death protein from carnation flower. Biochemistry 44:16377-84
Liu, Sijiu; Lu, Zhibing; Han, Yin et al. (2005) Crystal structures of 2-methylisocitrate lyase in complex with product and with isocitrate inhibitor provide insight into lyase substrate specificity, catalysis and evolution. Biochemistry 44:2949-62
Liu, Sijiu; Lu, Zhibing; Han, Ying et al. (2004) Conformational flexibility of PEP mutase. Biochemistry 43:4447-53
Herzberg, Osnat; Chen, Celia C H; Liu, Sijiu et al. (2002) Pyruvate site of pyruvate phosphate dikinase: crystal structure of the enzyme-phosphonopyruvate complex, and mutant analysis. Biochemistry 41:780-7

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