title: Sesquiterpene lactones inhibit luciferase but not beta-galactosidase activity in vitro and ex vivo.
abstract: Reporter enzymes such as firefly luciferase or beta-galactosidase of Escherichia coli are frequently used to study transcriptional activity of genes and to investigate the effects of novel compounds on gene or transcription factor activity. It is generally assumed that the activity of these enzymes is unaffected by the treatment conditions. Therefore, this factor is not considered when interpreting the data obtained. Biologically active compounds such as sesquiterpene lactones (SLs) have also been tested in reporter gene assays for their influence on gene expression. Here we show in in vitro and ex vivo experiments that SLs inhibit firefly luciferase activity probably by direct targeting of the enzyme while beta-galactosidase remains almost completely unaffected. The loss of luciferase activity after SL treatment could be an effect of their sulfhydryl-modifying potency and the subsequent alteration of the enzyme's tertiary structure. These results demonstrate that the effect of the test substance on the reporter enzyme used should be taken into consideration when the transcriptional effect of novel compounds is investigated.
authors.affiliation: Institut für Pharmazeutische Wissenschaften, Lehrstuhl für Pharmazeutische Biologie, Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Str. 19, 79104 Freiburg, Germany.
authors.forename: Maja T
authors.initials: MT
authors.lastname: Lindenmeyer
authors.forename: Alfonso J
authors.initials: AJ
authors.lastname: García-Piñeres
authors.forename: Victor
authors.initials: V
authors.lastname: Castro
authors.forename: Irmgard
authors.initials: I
authors.lastname: Merfort
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D004926:Escherichia coli / Q000201:enzymology; D017930:Genes, Reporter; D007783:Lactones / Q000737:chemistry / Q000494:pharmacology*; D008156:Luciferases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000235:genetics; D012717:Sesquiterpenes / Q000737:chemistry / Q000494:pharmacology*; D013223:Statistics as Topic; D013997:Time Factors; D014158:Transcription, Genetic / Q000187:drug effects; D001616:beta-Galactosidase / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D007783:Lactones; D016328:NF-kappa B; D012717:Sesquiterpenes; D008156:Luciferases; D001616:beta-Galactosidase