This project proposes the study of the structure, detailed function, and significance of the enzyme folylpolyglutamate synthetase to the chemotherapy of human tumors with folate antimetabolites. The research to be pursued entails a combination of biochemical, molecular and structural studies aimed at understanding how the enzyme converts folylmonoglutamates successively to oligo and polyglutamates. The shape of the active site will be examined and the distribution of hydrophobic and charged residues within the active site. The bioorganic chemistry of the reaction will be investigated using recombinant human enzyme and a series of probes of the active site. The structural differences apparent between the enzyme expressed in some tissues will be determined as will the molecular mechanisms responsible for control of expression of the mRNA for these species. A convergent series of techniques will be applied from expression of recombinant enzymes and active site biochemical studies to transcriptional analysis to structure-based drug design.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
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Experimental Therapeutics Subcommittee 1 (ET)
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Forry, Suzanne L
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Virginia Commonwealth University
Schools of Medicine
United States
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Yang, Chen; Xie, Lin-Ying; Windle, Jolene J et al. (2014) Humanizing mouse folate metabolism: conversion of the dual-promoter mouse folylpolyglutamate synthetase gene to the human single-promoter structure. FASEB J 28:1998-2008
Lawrence, Scott A; Titus, Steven A; Ferguson, Jennifer et al. (2014) Mammalian mitochondrial and cytosolic folylpolyglutamate synthetase maintain the subcellular compartmentalization of folates. J Biol Chem 289:29386-96
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Racanelli, Alexandra C; Turner, Fiona B; Xie, Lin-Ying et al. (2008) A mouse gene that coordinates epigenetic controls and transcriptional interference to achieve tissue-specific expression. Mol Cell Biol 28:836-48
Tomsho, John W; Moran, Richard G; Coward, James K (2008) Concentration-dependent processivity of multiple glutamate ligations catalyzed by folylpoly-gamma-glutamate synthetase. Biochemistry 47:9040-50
Chattopadhyay, Shrikanta; Moran, Richard G; Goldman, I David (2007) Pemetrexed: biochemical and cellular pharmacology, mechanisms, and clinical applications. Mol Cancer Ther 6:404-17
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
Turner, F B; Taylor, S M; Moran, R G (2000) Expression patterns of the multiple transcripts from the folylpolyglutamate synthetase gene in human leukemias and normal differentiated tissues. J Biol Chem 275:35960-8
Zhao, R; Titus, S; Gao, F et al. (2000) Molecular analysis of murine leukemia cell lines resistant to 5, 10-dideazatetrahydrofolate identifies several amino acids critical to the function of folylpolyglutamate synthetase. J Biol Chem 275:26599-606

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