Abstract

Folypolyglutamate synthetase (FPGS) catalyzes the addition of glutamic acid to folates and antifolates and as a result, causes the retention of these compounds in normal and neoplastic cells. The overall goals of this research are to understand the mechanism of the reaction catalyzed by the enzyme isolated from neoplastic tissue, to study the turnover of the enzyme in growing, static and differentiating tissues and to evaluate gene transfer methods for cloning the gene that codes for mammalian FPGS. The proposed work would (1) purify FPGS from rodent and human tumors, (2) kinetically characterize the sequence of addition of substrates to these enzymes, (3) study the time course of expression of enzyme activity in developing and differentiating tissues, (4) investigate the turnover of enzyme, and (5) attempt the isolation of a DNA clone that codes for human enzyme. These studies offer a better understanding of how FPGS works, when it is expressed and how its synthesis is controlled.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA027605-07
Application #
3167731
Study Section
Experimental Therapeutics Subcommittee 2 (ET)
Project Start
1980-04-01
Project End
1989-06-30
Budget Start
1986-07-01
Budget End
1987-06-30
Support Year
7
Fiscal Year
1986
Total Cost
Indirect Cost

  Institution

Name
Children's Hospital of Los Angeles
Department
Type
DUNS #
094878337
City
Los Angeles
State
CA
Country
United States
Zip Code
90027

  Publications

Racanelli, Alexandra C; Rothbart, Scott B; Heyer, Cortney L et al. (2009) Therapeutics by cytotoxic metabolite accumulation: pemetrexed causes ZMP accumulation, AMPK activation, and mammalian target of rapamycin inhibition. Cancer Res 69:5467-74
McCarthy, Erin A; Titus, Steven A; Taylor, Shirley M et al. (2004) A mutation inactivating the mitochondrial inner membrane folate transporter creates a glycine requirement for survival of chinese hamster cells. J Biol Chem 279:33829-36
Bronder, Julie L; Moran, Richard G (2003) A defect in the p53 response pathway induced by de novo purine synthesis inhibition. J Biol Chem 278:48861-71
Bronder, Julie L; Moran, Richard G (2002) Antifolates targeting purine synthesis allow entry of tumor cells into S phase regardless of p53 function. Cancer Res 62:5236-41
Andreassi 2nd, John L; Moran, Richard G (2002) Mouse folylpoly-gamma-glutamate synthetase isoforms respond differently to feedback inhibition by folylpolyglutamate cofactors. Biochemistry 41:226-35
Roberts, J D; Poplin, E A; Tombes, M B et al. (2000) Weekly lometrexol with daily oral folic acid is appropriate for phase II evaluation. Cancer Chemother Pharmacol 45:103-10
Titus, S A; Moran, R G (2000) Retrovirally mediated complementation of the glyB phenotype. Cloning of a human gene encoding the carrier for entry of folates into mitochondria. J Biol Chem 275:36811-7
Moran, R G (1999) Roles of folylpoly-gamma-glutamate synthetase in therapeutics with tetrahydrofolate antimetabolites: an overview. Semin Oncol 26:24-32
Tse, A; Brigle, K; Taylor, S M et al. (1998) Mutations in the reduced folate carrier gene which confer dominant resistance to 5,10-dideazatetrahydrofolate. J Biol Chem 273:25953-60
Tse, A; Moran, R G (1998) Cellular folates prevent polyglutamation of 5, 10-dideazatetrahydrofolate. A novel mechanism of resistance to folate antimetabolites. J Biol Chem 273:25944-52

Showing the most recent 10 out of 33 publications