title: Functional artificial luciferases as an optical readout for bioassays.
abstract: This study elucidates functional artificial luciferases (ALucs) wholly synthesized for bioassays and molecular imaging. The ALucs bearing epitopes were newly created by amending the sequences of our previously reported ALucs in light of a multi-sequence alignment and hydrophobicity search. The synthesized ALucs are survived in live cells and stable in culture media for 25 days after secretion. The epitopes in ALucs are exposed during the secretion process and indeed valid for column purification and immunological assays. The ALucs exerted a 9400-times stronger optical intensity with a coelenterazine derivative (CTZ i), when compared with Renilla reniformis luciferase 8.6-535. A supersecondary structure of ALuc30 was predicted with respect to the X-ray crystallographic information of the coelenterazine-binding protein (CBP). The structure revealed that ALuc30 has a room for accommodating the iodide of CTZ i. This study guides on how to create functional artificial luciferases and predicts the structural details with the current bioinformatics technologies.
authors.affiliation: Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba 305-8569, Japan. Electronic address: kimu-sb@aist.go.jp.
authors.forename: Sung Bae
authors.initials: SB
authors.lastname: Kim
authors.affiliation: Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST), 16-1 Onogawa, Tsukuba 305-8569, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Izumi
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D040761:Biomimetic Materials / Q000138:chemical synthesis / Q000737:chemistry; D019556:COS Cells; D002522:Chlorocebus aethiops; D004795:Enzyme Stability; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D008156:Luciferases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D008163:Luminescent Measurements / Q000379:methods*; D008958:Models, Molecular; D057054:Molecular Imaging; D008969:Molecular Sequence Data; D055620:Optical Phenomena; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D013379:Substrate Specificity
chemical_list: D000939:Epitopes; D011994:Recombinant Proteins; D008156:Luciferases
keywords: Bioluminescence; Copepod; Epitope; Imaging; Luciferase