Evaluation of firefly and Renilla luciferase inhibition in reporter-gene assays : a case of isoflavonoids

Kenda, Maša (avtor), Vegelj, Jan (avtor), Herlah, Barbara (avtor), Perdih, Andrej (avtor), Mladěnka, Přemysl (avtor), Sollner Dolenc, Marija (avtor)

Firefly luciferase is susceptible to inhibition and stabilization by compounds under investigation for biological activity and toxicity. This can lead to false-positive results in in vitro cell-based assays. However, firefly luciferase remains one of the most commonly used reporter genes. Here, we evaluated isoflavonoids for inhibition of firefly luciferase. These natural compounds are often studied using luciferase reporter-gene assays. We used a quantitative structure–activity relationship (QSAR) model to compare the results of in silico predictions with a newly developed in vitro assay that enables concomitant detection of inhibition of firefly and Renilla luciferases. The QSAR model predicted a moderate to high likelihood of firefly luciferase inhibition for all of the 11 isoflavonoids investigated, and the in vitro assays confirmed this for seven of them: daidzein, genistein, glycitein, prunetin, biochanin A, calycosin, and formononetin. In contrast, none of the 11 isoflavonoids inhibited Renilla luciferase. Molecular docking calculations indicated that isoflavonoids interact favorably with the D-luciferin binding pocket of firefly luciferase. These data demonstrate the importance of reporter-enzyme inhibition when studying the effects of such compounds and suggest that this in vitro assay can be used to exclude false-positives due to firefly or Renilla luciferase inhibition, and to thus define the most appropriate reporter gene.

• Jezik: Angleški jezik
• Ključne besede: isoflavonoids, firefly luciferase, Renilla luciferase, inhibition, quantitative structure–activity relationships, molecular docking, reporter-gene assay, OECD test guidelines, alternative methods
• Vrsta gradiva: Članek v reviji
• Tipologija: 1.01 - Izvirni znanstveni članek
• Organizacija: FFA - Fakulteta za farmacijo
• Status publikacije: Objavljeno
• Različica publikacije: Objavljena publikacija
• Leto izida: 2021
• Št. strani: 14 str.
• Številčenje: Vol. 22, iss. 13, art. 6927
• PID: 20.500.12556/RUL-135754
• UDK: 547.972:615.4.:54
• ISSN pri članku: 1422-0067
• DOI: 10.3390/ijms22136927
• COBISS.SI-ID: 72671747
• Datum objave v RUL: 30.03.2022

Gradivo je del revije

• Naslov: International journal of molecular sciences
• Skrajšan naslov: Int. j. mol. sci.
• Založnik: MDPI
• ISSN: 1422-0067
• COBISS.SI-ID: 2779162

Licence

• Licenca: CC BY 4.0, Creative Commons Priznanje avtorstva 4.0 Mednarodna
• Povezava: http://creativecommons.org/licenses/by/4.0/deed.sl
• Opis: To je standardna licenca Creative Commons, ki daje uporabnikom največ možnosti za nadaljnjo uporabo dela, pri čemer morajo navesti avtorja.

Sekundarni jezik

• Jezik: Slovenski jezik
• Ključne besede: izoflavonoidi, zaviralci, alternativne metode, molekularno mapiranje, farmacevtska kemija, flavonoidi