Establishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators

Abstract Human GLUT5 is a fructose-specific transporter in the glucose transporter family (GLUT, SLC2 gene family). Its substrate-specificity and tissue-specific expression make it a promising target for treatment of diabetes, metabolic syndrome and cancer, but few GLUT5 inhibitors are known. To ide...

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Autores principales:	Joanna Tripp, Christine Essl, Cristina V. Iancu, Eckhard Boles, Jun-yong Choe, Mislav Oreb
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/304ac976992841c9828f6fc9c777528f
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spelling	oai:doaj.org-article:304ac976992841c9828f6fc9c777528f2021-12-02T15:06:10ZEstablishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators10.1038/s41598-017-06262-42045-2322https://doaj.org/article/304ac976992841c9828f6fc9c777528f2017-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-06262-4https://doaj.org/toc/2045-2322Abstract Human GLUT5 is a fructose-specific transporter in the glucose transporter family (GLUT, SLC2 gene family). Its substrate-specificity and tissue-specific expression make it a promising target for treatment of diabetes, metabolic syndrome and cancer, but few GLUT5 inhibitors are known. To identify and characterize potential GLUT5 ligands, we developed a whole-cell system based on a yeast strain deficient in fructose uptake, in which GLUT5 transport activity is associated with cell growth in fructose-based media or assayed by fructose uptake in whole cells. The former method is convenient for high-throughput screening of potential GLUT5 inhibitors and activators, while the latter enables detailed kinetic characterization of identified GLUT5 ligands. We show that functional expression of GLUT5 in yeast requires mutations at specific positions of the transporter sequence. The mutated proteins exhibit kinetic properties similar to the wild-type transporter and are inhibited by established GLUT5 inhibitors N-[4-(methylsulfonyl)-2-nitrophenyl]-1,3-benzodioxol-5-amine (MSNBA) and (−)-epicatechin-gallate (ECG). Thus, this system has the potential to greatly accelerate the discovery of compounds that modulate the fructose transport activity of GLUT5.Joanna TrippChristine EsslCristina V. IancuEckhard BolesJun-yong ChoeMislav OrebNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Joanna Tripp Christine Essl Cristina V. Iancu Eckhard Boles Jun-yong Choe Mislav Oreb Establishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators
description	Abstract Human GLUT5 is a fructose-specific transporter in the glucose transporter family (GLUT, SLC2 gene family). Its substrate-specificity and tissue-specific expression make it a promising target for treatment of diabetes, metabolic syndrome and cancer, but few GLUT5 inhibitors are known. To identify and characterize potential GLUT5 ligands, we developed a whole-cell system based on a yeast strain deficient in fructose uptake, in which GLUT5 transport activity is associated with cell growth in fructose-based media or assayed by fructose uptake in whole cells. The former method is convenient for high-throughput screening of potential GLUT5 inhibitors and activators, while the latter enables detailed kinetic characterization of identified GLUT5 ligands. We show that functional expression of GLUT5 in yeast requires mutations at specific positions of the transporter sequence. The mutated proteins exhibit kinetic properties similar to the wild-type transporter and are inhibited by established GLUT5 inhibitors N-[4-(methylsulfonyl)-2-nitrophenyl]-1,3-benzodioxol-5-amine (MSNBA) and (−)-epicatechin-gallate (ECG). Thus, this system has the potential to greatly accelerate the discovery of compounds that modulate the fructose transport activity of GLUT5.
format	article
author	Joanna Tripp Christine Essl Cristina V. Iancu Eckhard Boles Jun-yong Choe Mislav Oreb
author_facet	Joanna Tripp Christine Essl Cristina V. Iancu Eckhard Boles Jun-yong Choe Mislav Oreb
author_sort	Joanna Tripp
title	Establishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators
title_short	Establishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators
title_full	Establishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators
title_fullStr	Establishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators
title_full_unstemmed	Establishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators
title_sort	establishing a yeast-based screening system for discovery of human glut5 inhibitors and activators
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/304ac976992841c9828f6fc9c777528f
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Establishing a yeast-based screening system for discovery of human GLUT5 inhibitors and activators

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