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Pharmaceutics 2018, 10(4), 186; https://doi.org/10.3390/pharmaceutics10040186

Development of Novel Intramolecular FRET-Based ABC Transporter Biosensors to Identify New Substrates and Modulators

Department of Chemistry & Biochemistry, College of Arts and Sciences, South Dakota State University, Brookings, SD 57007, USA
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Received: 17 September 2018 / Revised: 10 October 2018 / Accepted: 12 October 2018 / Published: 13 October 2018
(This article belongs to the Special Issue ABC Transporter-Mediated Drug Disposition)
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Abstract

Multidrug resistance protein 1 (MRP1) can efflux a wide variety of molecules including toxic chemicals, drugs, and their derivatives out of cells. Substrates of MRP1 include anti-cancer agents, antibiotics, anti-virals, anti-human immunodeficiency virus (HIV), and many other drugs. To identify novel substrates and modulators of MRP1 by exploiting intramolecular fluorescence resonance energy transfer (FRET), we genetically engineered six different two-color MRP1 proteins by changing green fluorescent protein (GFP) insertion sites, while keeping the red fluorescent protein (RFP) at the C-terminal of MRP1. Four of six recombinant proteins showed normal expression, localization, and transport activity. We quantified intramolecular FRET using ensemble fluorescence spectroscopy in response to binding of known substrate or ATP alone, substrate/ATP, and trapping of the transporter in closed conformation by vanadate. Recombinant MRP1 proteins GR-881, GR-888, and GR-905 exhibited reproducible and higher FRET changes under all tested conditions and are very promising for use as MRP1 biosensors. Furthermore, we used GR-881 to screen 40 novel anti-cancer drugs and identified 10 hits that potentially directly interact with MRP1 and could be substrates or modulators. Profiling of drug libraries for interaction with MRP1 can provide very useful information to improve the efficacy and reduce the toxicity of various therapies. View Full-Text
Keywords: ATP-binding cassette (ABC) proteins; multidrug resistance; fluorescence resonance energy transfer; biosensors; multidrug resistance protein 1; two-color MRP1 ATP-binding cassette (ABC) proteins; multidrug resistance; fluorescence resonance energy transfer; biosensors; multidrug resistance protein 1; two-color MRP1
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Osa-Andrews, B.; Tan, K.W.; Sampson, A.; Iram, S.H. Development of Novel Intramolecular FRET-Based ABC Transporter Biosensors to Identify New Substrates and Modulators. Pharmaceutics 2018, 10, 186.

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