Date of Award

2024

Document Type

Open Access Master's Report

Degree Name

Master of Science in Chemistry (MS)

Administrative Home Department

Department of Chemistry

Advisor 1

Marina Tanasova

Committee Member 1

Athar Ata

Committee Member 2

Shiyue Fang

Abstract

The mammalian facilitative glucose transporter (GLUT) family comprises 14 members that mediate the transport of hexoses across cell membranes. GLUT5 is the only member specific to fructose, and together with GLUT2, which transports fructose in addition to glucose, they make up the primary fructose transporters in humans. This study introduces a novel two-color fluorescence assay designed to simultaneously monitor the activity of GLUT5 and GLUT2 in live cells. 2,5-anhydro-D-mannitol (2,5-AM) a GLUT5 targeting compound has facilitated the development of various fructose-mimicking probes with a wide range of properties, such as radioactive imaging, fluorescent, photoactive, and drug delivery capabilities. Effective and accurate screening of fructose transporters GLUT5 and GLUT2 is essential for understanding their unique roles in cellular metabolism and their implications in metabolic disorders. This assay employed two fluorescent probes: P2, a mimicked conformation and configuration variation of fructose, and NBDM, a glucose analog that serves as a GLUT2 substrate. By leveraging their distinct fluorescence properties, P2 and NBDM facilitate the simultaneous visualization of GLUT5 and GLUT2 activities and this assay was validated with triple-negative breast cancer (TNBC) (MDA MB 231, 453,), hepatocellular carcinomas cell line (HEP-G2) proving its ability to differentiate fructose uptake by GLUT5 and GLUT2. This dual-color fluorescence assay marks significant progress in transporter research and has the potential for accelerating drug discovery and elucidating the molecular mechanisms underlying related metabolic diseases.

Additional Files
00 Publishing Agreement 9-2021.pdf (77 kB)
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