title: Superluminescent variants of marine luciferases for bioassays.
abstract: In this study, a rational synthesis of superluminescent variants from marine luciferases with prolonged bioluminescence has been demonstrated. A putative active site of a model marine luciferase, Gaussia princeps Luciferase (GLuc), was assigned and modified by a site-directed mutagenesis. The potent variants were found to generate up to 10 times stronger bioluminescence, emitting red shifts of up to 33 nm with natural coelenterazine than native GLuc, rendering an efficient optical signature in bioassays. The advantageous properties were demonstrated with mammalian two-hybrid assays, single-chain probes, and metastases of murine B16 melanoma in BALB/c nude mice. The unique ideas for engineering GLuc are proved to be valid even for other marine luciferases.
authors.affiliation: Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), Onogawa, Tsukuba, Japan. kimu-sb@aist.go.jp
authors.forename: Sung Bae
authors.initials: SB
authors.lastname: Kim
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Suzuki
authors.forename: Moritoshi
authors.initials: M
authors.lastname: Sato
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Tao
mesh_terms: D000818:Animals; D001681:Biological Assay; D019556:COS Cells; D002522:Chlorocebus aethiops; D003445:Crustacea / Q000201:enzymology; D014644:Genetic Variation / Q000235:genetics*; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D049449:Luminescence; D008163:Luminescent Measurements; D051379:Mice; D016296:Mutagenesis
chemical_list: D008156:Luciferases