Renilla Reniformis luciferase structure

The 3D structure of Renilla reniformis luciferase (RLuc) is solved. The tertiary structure was succesfully determined from high resolution (1.4 A) crystallographic analysis as illustrated in the volume 378 of the Journal of Molecular Biology by the group of Gambhir at the Stanford Molecular Imaging Program. Since the cloning of the gene for RLuc in 1991, this kind of luciferase from Sea pansy has been widely used in molecular biology, mainly to normalize reporter gene assays. Indeed, the widest application consists in the dual luciferase assay (DLA) in which the Renilla (driven by a basal promoter) normalizes the output of Firefly luciferase (tested promoter). Recently, work emerged also utilizing Rluc for in vivo imaging, to create novel imaging probes by fusing the luciferase to engineered antibodies, and to generate self-illuminating quantum dots. In this blog, protein interaction studies using Renilla were previously reviewed.

Now, structural data freely available in the protein data bank site (PDB) can help scientists to find more effective way to employ the luciferase (i.e., screening potential steric indrances prior to the creation of fusion protein constructs). Moreover, the crystallographic model (a classic alpha/beta hydrolase fold) confirms many amino-acid residues that are important for spectral and luminescence properties. In the same paper, the structure of the Renilla GFP (RrGFP), that together with the luciferase is responsible for the green light in the Sea pansy (via resonance energy transfer), has been also described.