A convenient, high-throughput assay for measuring the relative cell permeability of synthetic compounds.
We describe here a convenient procedure for assessing the relative cell permeability of chemical compounds. The assay can be used in a high-throughput mode and is particularly applicable for the evaluation of the relative permeability of compounds in a combinatorial library. The compound of interest is conjugated to a dexamethasone derivative. The entry of the conjugate into living mammalian cells triggers the nuclear transport of a Gal4 DNA binding domain-glucocorticoid receptor ligand binding domain-VP16 activation domain fusion protein and, consequently, the activation of a Gal4-responsive luciferase reporter gene. The relative cell permeability is thus described quantitatively by the level of luciferase expression. The experiments take only a few days once chemical synthesis and conjugation are finished.
Yu
Peng
P
Division of Translational Research, Departments of Internal Medicine and Molecular Biology, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, TX 75390-9185, USA.
Liu
Bo
B
Kodadek
Thomas
T
D002463:Cell Membrane Permeability / Q000502:physiology*; D020650:Combinatorial Chemistry Techniques; D003584:Cytological Techniques / Q000379:methods; D004268:DNA-Binding Proteins; D003907:Dexamethasone; D004789:Enzyme Activation / Q000187:drug effects; D008156:Luciferases / Q000378:metabolism; D009930:Organic Chemicals / Q000378:metabolism* / Q000494:pharmacology; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D009930:Organic Chemicals; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D003907:Dexamethasone; D008156:Luciferases