eUnaG: a new ligand-inducible fluorescent reporter to detect drug transporter activity in live cells.
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.
Department of Genetics, Harvard Medical School, Boston, MA, 02115, USA.
Johannes T-H
JT
Yeh
Department of Chemistry and Biochemistry, University of Texas at Arlington, Arlington, TX, 76019-0065, USA.|Department of Chemistry, Umeå University, Umeå, SE-901 87, Sweden.
Kwangho
K
Nam
Institute of Mathematics, National Taiwan University, Taipei, 10617 Taiwan.
Joshua T-H
JT
Yeh
Department of Genetics, Harvard Medical School, Boston, MA, 02115, USA.|Howard Hughes Medical Institute, Harvard Medical School, Boston, MA, 02115, USA.
Norbert
N
Perrimon
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001663:Bilirubin / Q000378:metabolism*; D001692:Biological Transport; D015374:Biosensing Techniques; D004330:Drosophila; D055808:Drug Discovery; D015870:Gene Expression; D017930:Genes, Reporter; D006801:Humans; D008024:Ligands; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D008958:Models, Molecular; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013312:Stress, Physiological; D013329:Structure-Activity Relationship
D018528:ATP-Binding Cassette Transporters; D008024:Ligands; D026901:Membrane Transport Proteins; D011993:Recombinant Fusion Proteins; D001663:Bilirubin