title: Development of luciferin analogues bearing an amino group and their application as BRET donors.
abstract: We systematically synthesized bioluminogenic substrates bearing an amino group on benzothiazole, quinoline, naphthalene, and coumarin scaffolds. They emit bioluminescence in various colors: red, orange, yellow, and green. An amino-substituted coumarylluciferin derivative, coumarylaminoluciferin (CAL), showed the shortest bioluminescence wavelength among substrates reported so far. Further, the fluorescence of CAL did not exhibit solvatochromism, which suggests that its bioluminescence is not susceptible to environmental factors. We applied CAL as an energy-donor substrate for a bioluminescence resonance energy transfer (BRET) system with click beetle red luciferase (CBRluc), a mutant of firefly luciferase, as the energy-donor enzyme and yellow fluorescent protein (YFP) as the energy-acceptor fluorophore, and obtained a clearly bimodal bioluminescence spectrum. Stable bioluminescence that is not influenced by environmental factors is highly desirable for reliable measurements in biological assays.
authors.affiliation: Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
authors.forename: Hideo
authors.initials: H
authors.lastname: Takakura
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Sasakura
authors.forename: Tasuku
authors.initials: T
authors.lastname: Ueno
authors.forename: Yasuteru
authors.initials: Y
authors.lastname: Urano
authors.forename: Takuya
authors.initials: T
authors.lastname: Terai
authors.forename: Kenjiro
authors.initials: K
authors.lastname: Hanaoka
authors.forename: Takashi
authors.initials: T
authors.lastname: Tsuboi
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nagano
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D052160:Benzothiazoles / Q000737:chemistry; D003374:Coumarins / Q000737:chemistry; D049454:Firefly Luciferin / Q000031:analogs & derivatives* / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D008156:Luciferases / Q000378:metabolism; D049408:Luminescent Agents / Q000737:chemistry*; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000737:chemistry; D009281:Naphthalenes / Q000737:chemistry; D011804:Quinolines / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D052160:Benzothiazoles; D003374:Coumarins; D049408:Luminescent Agents; D008164:Luminescent Proteins; D009281:Naphthalenes; D011804:Quinolines; C054562:yellow fluorescent protein, Bacteria; C031721:naphthalene; D049454:Firefly Luciferin; C030123:coumarin; C037219:quinoline; D008156:Luciferases; C005465:benzothiazole