title: The role of firefly luciferase C-terminal domain in efficient coupling of adenylation and oxidative steps.
abstract: The N-terminal domain (N-domain) of the firefly luciferase from Photinus pyraris has weak luminescence activity, and shows a unique light emitting profile with very long rise time of more than several minutes. Through a sensitive assay of the reaction intermediate luciferyl-adenylate (LH2-AMP), we found that the slow increase in the N-domain luminescence faithfully reflected the concentration of dissociated LH2-AMP. No such correlation was observed for wild-type or mutant enzymes with short rise time, except one with longer rise time. The results suggest that the C-terminal domain plays an indispensable role in efficiently coupling adenylation and oxidative steps.
authors.affiliation: Department of Chemistry and Biotechnology, School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Keiichi
authors.initials: K
authors.lastname: Ayabe
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Zako
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ueda
mesh_terms: D000249:Adenosine Monophosphate / Q000378:metabolism; D000818:Animals; D048888:Fireflies / Q000201:enzymology; D049409:Luciferases, Firefly / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D010084:Oxidation-Reduction; D017434:Protein Structure, Tertiary / Q000235:genetics / Q000502:physiology; D013379:Substrate Specificity
chemical_list: D000249:Adenosine Monophosphate; D049409:Luciferases, Firefly