Luciferase from Fulgeochlizus bruchi (Coleoptera:Elateridae), a Brazilian click-beetle with a single abdominal lantern: molecular evolution, biological function and comparison with other click-beetle luciferases

Abstract

Bioluminescent click-beetles emit a wide range of bioluminescence colors (λ(Max) = 534-594 nm) from thoracic and abdominal lanterns, which are used for courtship. Only the luciferases from Pyrophorus and Pyrearinus species were cloned and sequenced. The Brazilian Fulgeochlizus bruchi click-beetle, which inhabits the Central-west Cerrado (Savannas), is noteworthy because, differently from other click-beetles, the adult stage displays only a functional abdominal lantern, which produces a bright green bioluminescence for sexual attraction purposes, and lacks functional thoracic lanterns. We cloned the cDNA for the abdominal lantern luciferase of this species. Notably, the primary sequence of this luciferase showed slightly higher identity with the green emitting dorsal lantern luciferases of the Pyrophorus genus instead of the abdominal lanterns luciferases. This luciferase displays a blue-shifted spectrum (λ(Max) = 540 nm), which is pH-insensitive from pH 7.5 to 9.5 and undergoes a slight red shift and broadening above this pH; the lowest K(M) for luciferin among studied click-beetle luciferases, and the highest optimum pH (9.0) ever reported for a beetle luciferase. At pH 9.0, the K(M) for luciferin increases, showing a decrease of affinity for this substrate, despite the higher activity. The slow luminescence decay rate of F. bruchi luciferase in vitro reaction could be an adaptation of this luciferase for the long and sustained in vivo luminescence display of the click-beetle during the courtship, and could be useful for in vivo intracellular imaging.

Molecular phylogeny of Neotropical bioluminescent beetles (Coleoptera: Elateroidea) in southern and central Brazil

“…Our analyses also showed a closer relationship between the species Pyrophorus divergens and Fulgeochlizus bruchi . A similar result was also shown by luciferases sequences analysis, with high support value , and by morphologic studies . However, only a few genera were used in this analysis, and other taxa are necessary for more detailed analysis.…”

“…Some bionomic studies on Brazilian species were carried out for the Lampyridae and Phengodidae . Biochemical and molecular evolution analyses were carried out for the luciferases of the Brazilian click beetles Pyrearinus termitilluminans , Fulgeochlizus bruchi , Mastinocerini railroadworms of the genera Phrixothrix and Brasilocerus and fireflies of the genera Cratomorphus , Macrolampis and Amydetes .…”

Use of a special Brazilian red-light emitting railroad worm Luciferase in bioassays of NEK7 protein Kinase and Creatine Kinase

“…One of the most important sources of luciferases are the bioluminescent beetles belonging to the Elateroidea superfamily, within Lampyridae (fireflies), Phengodidae (railroadworms) and Elateridae (click-beetles) [1, 2]. They emit different bioluminescence colors depending on life stage and the lantern type, from green to red, that serve as ecological adaptations to different photic environments and for diverse biological functions.…”

The Effect of Surface Charge Saturation on Heat‐induced Aggregation of Firefly Luciferase

“…It has been reported that firefly luciferases show bright bioluminescence, low background signal, high catalytic efficiency, substrate specificity and high sensitivity to ATP, however, actual and potential applications of wild‐type beetle luciferases are often limited by insufficient stability or decrease in activity of the enzyme in a particular assay . It also has been mentioned that many novel beetle luciferases with promising properties successfully solve or alleviate many of these limitations. Different strategies have been applied such as site‐directed mutagenesis in thermolabile regions , to achieve more favorable luciferases to decrease sensitivity of this enzyme at higher temperatures.…”