RRC ID 72272
Author Kim SB, Torimura M, Tao H.
Title Creation of artificial luciferases for bioassays.
Journal Bioconjug Chem
Abstract The present study demonstrates the creation of artificial luciferases (ALuc) for bioassays, inspired by a sequence alignment of copepod luciferases. Extraction of the consensus amino acids from the alignment enabled us to generate a series of ALucs with unique optical properties and sequential identities that are clearly different from those of any existing copepod luciferase. For example, some ALucs exhibited heat stability, dramatically prolonged optical intensities, broad full width at half-maximum, and strong optical intensities. The practical suitability of the luciferases as an optical readout was examined in diverse bioassays, including mammalian two-hybrid assays, live cell imaging, single-chain probes, bioluminescent capsules, and bioluminescent antibodies. We further determine the physical properties of ALucs through bioinformatic analysis and finally discuss detailed issues on the unique properties of ALucs. The present study shows how to create the artificial enzymes with excellent optical properties for bioassays and encourages researchers to fabricate their own unique artificial enzymes with designed properties and functionalities.
Volume 24(12)
Pages 2067-75
Published 2013-12-18
DOI 10.1021/bc400411h
PMID 24237362
MeSH Animals Biological Assay / methods* Biomimetic Materials / chemistry Biomimetic Materials / metabolism* COS Cells Cell Survival Chlorocebus aethiops Computational Biology Copepoda / enzymology Enzyme Stability Hydrocortisone / metabolism Luciferases / chemistry Luciferases / metabolism* Mice Molecular Imaging Optical Phenomena
Resource
DNA material ALuc23 (RDB19616)