Abstract
The absorption, distribution, metabolism and excretion (ADME) of metabolites and toxic organic solutes are orchestrated by the ATP-binding cassette (ABC) transporters and the organic solute carrier family (SLC) proteins. A large number of ABC and SLC transpoters exist; however, only a small number have been well characterized. To facilitate the analysis of these transporters, which is important for drug safety and physiological studies, we developed a sensitive genetically encoded bilirubin (BR)-inducible fluorescence sensor (eUnaG) to detect transporter-coupled influx/efflux of organic compounds. This sensor can be used in live cells to measure transporter activity, as excretion of BR depends on ABC and SLC transporters. Applying eUnaG in functional RNAi screens, we characterize l(2)03659 as a Drosophila multidrug resistant-associated ABC transporter.
Highlights
The sensor we engineered is based on the genetically encoded bilirubin (BR)-inducible fluorescence protein UnaG
Excretion of BR is mostly mediated by the ABC transporters MRP1-4 and MDR, as well as the solute carrier family (SLC) transporters OATP1A (Slco1A) and OATP1B (Slco1B)
Based on the observation that MRPs/MDR ABC transporters and OATP transporters recognize a broad spectrum of substrates[7,15], including BR, bile acids and most drug compounds, we reasoned that UnaG could be used as a sensor for rapid assays to reflect the overall activities of ABC and OATP transporters in live cells
Summary
The sensor we engineered is based on the genetically encoded bilirubin (BR)-inducible fluorescence protein UnaG. One of the most abundant organic molecules in the body, is involved in oxygen carriage in red blood cells and acts as a co-factor for many enzymes such as the large detoxifiying P450 cytochrome family in the liver[14]. BR itself is abundant and needs to be efficiently removed from the body as it is toxic to cells. Based on the observation that MRPs/MDR ABC transporters and OATP transporters recognize a broad spectrum of substrates[7,15], including BR, bile acids and most drug compounds, we reasoned that UnaG could be used as a sensor for rapid assays to reflect the overall activities of ABC and OATP transporters in live cells. We decided to engineer a form of UnaG that could be used as a probe for measuring transporter activities in live cells
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
