Evaluation of Firefly and Renilla Luciferase Inhibition in Reporter-Gene Assays: A Case of Isoflavonoids

Abstract

Firefly luciferase remains one of the most commonly used reporter genes, but some compounds can interfere with this assay. The quantitative structure-activity relationship 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.

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 different compounds.

Keywords

• isoflavonoids
• firefly luciferase
• Renilla luciferase
• inhibition
• quantitative structure-activity relationships
• molecular docking
• reporter-gene assay
• OECD test guidelines
• alternative methods