Identification of novel small-molecule inhibitors for human transketolase by high-throughput screening with fluorescent intensity (FLINT) assay

Journal of Biomolecular Screening
Mark X DuBrad Carte

Abstract

The metabolic enzyme transketolase (TK) plays a crucial role in tumor cell nucleic acid synthesis, using glucose through the elevated nonoxidative pentose phosphate pathway (PPP). Identification of inhibitors specifically targeting TK and preventing the nonoxidative PPP from generating the RNA ribose precursor, ribose-5-phosphate, provides a novel approach for developing effective anticancer therapeutic agents. The full-length human transketolase gene was cloned and expressed in Escherichia coli and the recombinant human transketolase protein purified to homogeneity. A fluorescent intensity (FLINT) assay was developed and optimized. Library compounds were screened in a high-throughput screening (HTS) campaign using the FLINT assay. Fifty-four initial hits were identified. Among them, 2 scaffolds with high selectivity, ideal physiochemical properties, and low molecular weight were selected for lead optimization studies. These compounds specifically inhibited in vitro TK enzyme activity and suppressed tumor cell proliferation in at least 3 cancer cell lines: SW620, LS174T, and MIA PaCa-2. Identification of these active scaffolds represents a good starting point for development of drugs specifically targeting TK and the nonoxidati...Continue Reading

References

Dec 1, 1969·Journal of Bacteriology·B L Josephson, D G Fraenkel
Jan 1, 1982·Methods in Enzymology·G A Kochetov
Jan 22, 1998·Proceedings of the National Academy of Sciences of the United States of America·D C MacallanM K Hellerstein
Jun 5, 1998·The International Journal of Biochemistry & Cell Biology·G SchenkP F Nixon
Dec 19, 1998·Cell Biochemistry and Function·R M BabiakM B Oliveira
Jan 30, 1999·The International Journal of Biochemistry & Cell Biology·G SchenkP F Nixon
Jan 6, 2000·FEBS Letters·Olga N Solovjeva, G A Kochetov
Jun 6, 2000·Journal of Biomolecular Screening·J H ZhangK R Oldenburg

Citations

Jun 5, 2012·Ophthalmology·Clemens A LangeClaudia Auw-Haedrich
May 15, 2012·Free Radical Biology & Medicine·Chiara RigantiDario Ghigo
Jun 12, 2010·Molecular and Cellular Endocrinology·Bogusz TrojanowiczCuong Hoang-Vu
Sep 12, 2008·Bioorganic & Medicinal Chemistry Letters·Jialin MaoAlan P Kozikowski
Sep 25, 2012·Chembiochem : a European Journal of Chemical Biology·Dong YiWolf-Dieter Fessner
Jun 3, 2015·Chembiochem : a European Journal of Chemical Biology·Maria Chiara ForchinDaniela Monti
Jan 10, 2008·Bioorganic & Medicinal Chemistry Letters·Allen A ThomasBarb Brandhuber
Feb 13, 2008·Bioorganic & Medicinal Chemistry Letters·Allen A ThomasGuy Vigers
Dec 2, 2005·Human Molecular Genetics·Sakari VanharantaLauri A Aaltonen
Apr 26, 2017·Clinical Epigenetics·Sheraz Gul
Aug 25, 2016·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Kunitoshi ShigeyasuAjay Goel

Related Concepts

Enzyme Inhibitors
Fluorescence
Apotransketolase
Cell Line, Tumor
Cell Proliferation

Related Feeds

Cancer Metabolism

In order for cancer cells to maintain rapid, uncontrolled cell proliferation, they must acquire a source of energy. Cancer cells acquire metabolic energy from their surrounding environment and utilize the host cell nutrients to do so. Here is the latest research on cancer metabolism.