Department of Chemistry, UC Davis, One Shields Avenue, Davis, CA, United States.
Luminescence. 2023 Feb;38(2):216-220. doi: 10.1002/bio.4415. Epub 2022 Dec 4.
The development of bioluminescence-based tools has seen steady growth in the field of chemical biology over the past few decades ranging in uses from reporter genes to assay development and targeted imaging. More recently, coelenterazine-utilizing luciferases such as Gaussia, Renilla, and the engineered nano-luciferases have been utilized due to their intense luminescence relative to firefly luciferin/luciferase. The emerging importance of these systems warrants investigations into the components that affect their light production. Previous work has reported that one marine luciferase, Gaussia, is potently inhibited by copper salt. The mechanism for inhibition was not elucidated but was hypothesized to occur via binding to the enzyme. In this study, we provide the first report of a group of nonhomologous marine luciferases also exhibiting marked decreases in light emission in the presence of copper (II). We investigate the mechanism of action behind this inhibition and demonstrate that the observed copper inhibition does not stem from a luciferase interaction but rather the chemical oxidation of imidazopyrazinone luciferins generating inert, dehydrated luciferins.
在过去的几十年中，生物发光为基础的工具在化学生物学领域得到了稳步发展，其用途从报告基因到检测开发和靶向成像。最近，由于与萤火虫荧光素/荧光酶相比具有强烈的发光，海肾荧光素酶、Gaussia 荧光素酶和工程化的纳米荧光素酶等腔肠素利用型荧光素酶已被广泛应用。这些系统的重要性日益凸显，需要对影响其发光的因素进行研究。以前的研究报告表明，一种海洋荧光素酶 Gaussia 被铜盐强烈抑制。虽然抑制机制尚未阐明，但据推测是通过与酶结合而发生的。在这项研究中，我们首次报道了一组非同源海洋荧光素酶，它们在存在铜（II）的情况下也表现出明显的发光减少。我们研究了这种抑制作用的作用机制，并证明观察到的铜抑制不是源于荧光素酶的相互作用，而是由于imidazopyrazinone 荧光素的化学氧化生成惰性、脱水的荧光素。
