Isolation and characterization of the transient, luciferase-bound flavin-4a-hydroxide in the bacterial luciferase reaction

Kurfürst, Manfred; Macheroux, Peter; Ghisla, Sandro; Hastings, J. Woodland

Isolation and characterization of the transient, luciferase-bound flavin-4a-hydroxide in the bacterial luciferase reaction

Files

ghislascan154.pdf(331.04 KB)

Date

1987

Authors

Kurfürst, Manfred

Macheroux, Peter

Ghisla, Sandro

Hastings, J. Woodland

Publication type

Journal article

Published in

Biochimica et Biophysica Acta ; 924 (1987), 1. - pp. 104-110. - ISSN 0304-4165

Abstract

Procedures and conditions have been established such that the unstable enzyme-bound flavin intermediate produced in the bacterial luciferase reaction can be isolated as approximately 70% of the flavin product, the remaining being the final product, FMN. The structure of the intermediate is proposed to be that of a luciferase-bound 4a,5-dihydroflavin-4a-hydroxide. The intermediate has a half-life of 33 min at 2°C and decays spontaneously to give H2O and luciferase-bound FMN with an activation enthalpy of about 120 kJ/mol. It has an absorption spectrum (λmax = 360 nm) that is consistent with the proposed structure, and a fluorescence emission (λmax = 485 nm) that matches the bioluminescence emission closely.

Subject (DDC)

570 Biosciences, Biology

Keywords

Bioluminescence,Oxygen activation,Flavin intermediate,Luciferase,Bacteria

Cite This

ISO 690

KURFÜRST, Manfred, Peter MACHEROUX, Sandro GHISLA, J. Woodland HASTINGS, 1987. Isolation and characterization of the transient, luciferase-bound flavin-4a-hydroxide in the bacterial luciferase reaction. In: Biochimica et Biophysica Acta. 924(1), pp. 104-110. ISSN 0304-4165. Available under: doi: 10.1016/0304-4165(87)90076-6

BibTex

@article{Kurfurst1987Isola-7258,
  year={1987},
  doi={10.1016/0304-4165(87)90076-6},
  title={Isolation and characterization of the transient, luciferase-bound flavin-4a-hydroxide in the bacterial luciferase reaction},
  number={1},
  volume={924},
  issn={0304-4165},
  journal={Biochimica et Biophysica Acta},
  pages={104--110},
  author={Kurfürst, Manfred and Macheroux, Peter and Ghisla, Sandro and Hastings, J. Woodland}
}

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