title: Aminoluciferins extend firefly luciferase bioluminescence into the near-infrared and can be preferred substrates over D-luciferin.
abstract: Firefly luciferase adenylates and oxidizes d-luciferin to chemically generate visible light and is widely used for biological assays and imaging. Here we show that both luciferase and luciferin can be reengineered to extend the scope of this light-emitting reaction. D-Luciferin can be replaced by synthetic luciferin analogues that increase near-infrared photon flux >10-fold over that of D-luciferin in live luciferase-expressing cells. Firefly luciferase can be mutated to accept and utilize rigid aminoluciferins with high activity in both live and lysed cells yet exhibit 10,000-fold selectivity over the natural luciferase substrate. These new luciferin analogues thus pave the way to an extended family of bioluminescent reporters.
authors.affiliation: Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School , 364 Plantation Street, Worcester, Massachusetts 01605, United States.
authors.forename: David M
authors.initials: DM
authors.lastname: Mofford
authors.forename: Gadarla Randheer
authors.initials: GR
authors.lastname: Reddy
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Miller
mesh_terms: D000586:Amination; D000818:Animals; D052160:Benzothiazoles / Q000032:analysis / Q000138:chemical synthesis / Q000378:metabolism*; D016466:CHO Cells; D003412:Cricetulus; D048888:Fireflies / Q000201:enzymology; D007700:Kinetics; D049409:Luciferases, Firefly / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D049408:Luminescent Agents / Q000032:analysis / Q000138:chemical synthesis / Q000378:metabolism*; D008163:Luminescent Measurements; D009154:Mutation; D013379:Substrate Specificity
chemical_list: D052160:Benzothiazoles; C532924:D-luciferin; D049408:Luminescent Agents; D049409:Luciferases, Firefly