Abstract

This proposal outlines a research program whose goal is to determine the catalytic mechanism of the folate- dependent de novo purine biosynthetic enzyme, glycinamide ribonucleotide transformylase. A series of kinetic studies, substrate specificity studies, affinity labeling studies, and site-directed mutagenesis studies are described.
The aim of these studies is to define the kinetic and chemical mechanism of the transformylation reaction and to identify the enzyme residues involved in substrate binding and catalysis. These studies will afford mechanistic and structural information about the transformylase which will provide a basis for the design of specific transformylase inhibitors. These studies will also serve as the foundation for future studies on the interactions of the enzymes which constitute the de novo purine biosynthetic pathway and the mechanism(s) of regulation of the pathway. Although it has been demonstrated that inhibition of this pathway shows potential for cancer chemotherapy, much basic information about the pathway and the individual enzymes which constitute the pathway remains to be elucidated.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM042663-03
Application #
2181568
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Project Start
1992-09-30
Project End
1996-08-31
Budget Start
1994-09-01
Budget End
1995-08-31
Support Year
3
Fiscal Year
1994
Total Cost
Indirect Cost

  Institution

Name
University of Cincinnati
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
City
Cincinnati
State
OH
Country
United States
Zip Code
45221

  Publications

Dahms, Tanya E S; Sainz, Germaine; Giroux, Eugene L et al. (2005) The apo and ternary complex structures of a chemotherapeutic target: human glycinamide ribonucleotide transformylase. Biochemistry 44:9841-50
Antle, V D; Donat, N; Hua, M et al. (1999) Substrate specificity of human glycinamide ribonucleotide transformylase. Arch Biochem Biophys 370:231-5
Poch, M T; Qin, W; Caperelli, C A (1998) The human trifunctional enzyme of de novo purine biosynthesis: heterologous expression, purification, and preliminary characterization. Protein Expr Purif 12:17-24
Caperelli, C A; Giroux, E L (1997) The human glycinamide ribonucleotide transformylase domain: purification, characterization, and kinetic mechanism. Arch Biochem Biophys 341:98-103
Antle, V D; Liu, D; McKellars, B R et al. (1996) Substrate specificity of glycinamide ribonucleotide transformylase from chicken liver. J Biol Chem 271:6045-9