A Novel Multiple-Read Screen for Metabolically Active Compounds Based on a Genetically Encoded FRET Sensor for ATP.

Zhao, Ziyan; Rajagopalan, Rahul; Zweifach, Adam

Zhao, Ziyan; Rajagopalan, Rahul; Zweifach, Adam.

Afiliación

Zhao Z; 1 Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT, USA.
Rajagopalan R; 1 Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT, USA.
Zweifach A; 1 Department of Molecular and Cell Biology, University of Connecticut, Storrs, CT, USA.

SLAS Discov ; 23(9): 907-918, 2018 10.

Article en En | MEDLINE | ID: mdl-29898642

RESUMEN

Both glycolysis and mitochondrial energetics are targets of interest for developing antiproliferative cancer therapeutics. We developed a novel multiple-read assay based on long-term expression in K562 cells of a genetically encoded intramolecular Förster resonance energy transfer sensor for adenosine triphosphate (ATP). The assay, conducted in a fluorescent plate reader, can identify compounds that inhibit oxidative phosphorylation-dependent ATP production, glycolysis, or both after short-term treatment. We screened a National Cancer Institute (NCI) compound library, identifying inhibitors of oxidative phosphorylation-dependent ATP production and glycolysis. Three glycolysis inhibitors blocked hexokinase activity, demonstrating that our assay can serve as the initial step in a workflow to identify compounds that inhibit glycolysis via a defined desired mechanism. Finally, upon reviewing the literature, we found surprisingly little evidence that inhibiting glycolysis with small molecules is antiproliferative. Using NCI data on proliferation of K562 cells, we found that inhibitors of oxidative phosphorylation-dependent ATP production were no more antiproliferative than the overall library, whereas all glycolysis inhibitors were in the top third of most effective antiproliferative compounds. Our results thus present a powerful new way to screen for compounds that affect cellular metabolism and also provide important support for the idea that blocking glycosis is antiproliferative.

Asunto(s)

Adenosina Trifosfato/metabolismo; Técnicas Biosensibles; Descubrimiento de Drogas; Metabolismo Energético/efectos de los fármacos; Transferencia Resonante de Energía de Fluorescencia; Ingeniería Genética; Antineoplásicos/farmacología; Línea Celular Tumoral; Descubrimiento de Drogas/métodos; Transferencia Resonante de Energía de Fluorescencia/métodos; Ingeniería Genética/métodos; Glucólisis/efectos de los fármacos; Humanos; Células K562; Mitocondrias/efectos de los fármacos; Mitocondrias/metabolismo; Fosforilación Oxidativa/efectos de los fármacos; Fenotipo

Palabras clave

cancer therapeutics; cell proliferation; fluorescence resonance energy transfer (FRET); glycolysis; metabolism; oxidative phosphorylation

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Técnicas Biosensibles / Ingeniería Genética / Adenosina Trifosfato / Transferencia Resonante de Energía de Fluorescencia / Metabolismo Energético / Descubrimiento de Drogas Límite: Humans Idioma: En Revista: SLAS Discov Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos

Texto completo

Añadir a Mi BVS

Imprimir

XML

PubMed Links

Buscar en Google