Sesquiterpene lactones inhibit luciferase but not beta-galactosidase activity in vitro and ex vivo
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Sesquiterpene lactones inhibit luciferase but not beta-galactosidase activity in vitro and ex vivo. / Lindenmeyer, Maja T; García-Piñeres, Alfonso J; Castro, Victor; Merfort, Irmgard.
In: ANAL BIOCHEM, Vol. 328, No. 2, 15.05.2004, p. 147-54.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Sesquiterpene lactones inhibit luciferase but not beta-galactosidase activity in vitro and ex vivo
AU - Lindenmeyer, Maja T
AU - García-Piñeres, Alfonso J
AU - Castro, Victor
AU - Merfort, Irmgard
PY - 2004/5/15
Y1 - 2004/5/15
N2 - 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.
AB - 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.
KW - Animals
KW - Cells, Cultured
KW - Dose-Response Relationship, Drug
KW - Enzyme Inhibitors
KW - Escherichia coli
KW - Genes, Reporter
KW - Lactones
KW - Luciferases
KW - NF-kappa B
KW - Sesquiterpenes
KW - Statistics as Topic
KW - Time Factors
KW - Transcription, Genetic
KW - beta-Galactosidase
KW - Journal Article
KW - Research Support, Non-U.S. Gov't
U2 - 10.1016/j.ab.2004.01.021
DO - 10.1016/j.ab.2004.01.021
M3 - SCORING: Journal article
C2 - 15113690
VL - 328
SP - 147
EP - 154
JO - ANAL BIOCHEM
JF - ANAL BIOCHEM
SN - 0003-2697
IS - 2
ER -