MICROSCOPY & IMAGING
BRIGHT FUTURE FOR BIOLUMINESCENCE
Thomas Machleidt on exploring new applications for bioluminescence
O
ver the past three decades, bioluminescence has emerged as one of the most broadly used reporter technologies in life science
research. For the first two decades, the use of bioluminescence was largely dominated by ATP-dependent luciferase such as firefly luciferase. Te remarkable breadth of tools derived from this family of luciferases was predicated on its biophysical and chemical properties and can be divided in four principal assay categories: (a) transcriptional reporters for pathway analysis (b) viability detectors based on measurement of cellular ATP levels (c) pro-luciferins for analysis of enzyme activity (d) luciferase based biosensors for measuring physiological processes (Fig. 1). Despite the tremendous value firefly luciferase brought to life science, its utility proved limited by some of its intrinsic features, including the relatively low specific signal, large size, complex structure and ATP dependency.
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Fig. 1. Configurations of firefly luciferase To overcome these liabilities, Promega
developed a novel luciferase system called NanoLuc, which was originally derived from a bright, hetero-tetrameric luciferase isolated from the deep-sea shrimp Oplophorus gracilirostris. Within the enzyme complex, the 19kD sub-unit was found to be associated with luciferase activity. Tough rather dim
and unstable in its wildtype form, scientists were able to improve specific activity over two million-fold by synergistically combining directed evolution and substrate development. Te resulting luciferase produces a glow-type luminescence with a specific activity 100 times higher than firefly luciferase. Furthermore, the enzyme is small and thermostable up to
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