title: Quenching the firefly bioluminescence by various ions.
abstract: The luciferase reporter gene assay system is broadly applied in various biomedical aspects, including signaling pathway dissection, transcriptional activity analysis, and genetic toxicity testing. It significantly improves the experimental accuracy and reduces the experimental error by the addition of an internal control. In the current research, we discovered some specific ions that could selectively inhibit firefly luciferase while having a negligible effect on renilla luciferase in vitro in the dual-reporter gene assay. We showed that these ionic compounds had a high potential of being utilized as quench-and-activate reagents in the dual-reporter assay. Furthermore, results from kinetic studies on ion-mediated quenching effects indicated that different ions have distinct inhibition modes. Our study is anticipated to guide a more affordable design of quench-and-activate reagents in biomedicine and pharmaceutical analysis.
authors.affiliation: Department of Medicinal Chemistry, Key Laboratory of Chemical Biology (MOE), School of Pharmacy, Shandong University, Jinan, Shandong 250012, China. mli@sdu.edu.cn.
authors.forename: Huateng
authors.initials: H
authors.lastname: Zhang
authors.forename: Haixiu
authors.initials: H
authors.lastname: Bai
authors.forename: Tianyu
authors.initials: T
authors.lastname: Jiang
authors.forename: Zhao
authors.initials: Z
authors.lastname: Ma
authors.forename: Yanna
authors.initials: Y
authors.lastname: Cheng
authors.forename: Yubin
authors.initials: Y
authors.lastname: Zhou
authors.forename: Lupei
authors.initials: L
authors.lastname: Du
authors.forename: Minyong
authors.initials: M
authors.lastname: Li
mesh_terms: D000818:Animals; D057075:Enzyme Assays; D048888:Fireflies / Q000201:enzymology* / Q000235:genetics; D017930:Genes, Reporter; D007477:Ions / Q000378:metabolism*; D049409:Luciferases, Firefly / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D049410:Luciferases, Renilla / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D049449:Luminescence; D049408:Luminescent Agents / Q000378:metabolism*; D048790:Renilla / Q000201:enzymology* / Q000235:genetics
chemical_list: D007477:Ions; D049408:Luminescent Agents; D049410:Luciferases, Renilla; D049409:Luciferases, Firefly