Spectral detection of an intermediate preceding the excited state in the bacterial luciferase reaction

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

Spectral detection of an intermediate preceding the excited state in the bacterial luciferase reaction

Dateien

Spectral_detection_of_an_intermediate_preceding_the_excited_state_in_the_bacterial_luciferase_reaction.pdfGröße: 874.37 KBDownloads: 465

Datum

1993

Autor:innen

Macheroux, Peter

Ghisla, Sandro

Hastings, J. Woodland

Projekt

Biolumineszenz und Blaulichtwahrnehmung

Sammlungen

Biologie

Publikationstyp

Zeitschriftenartikel

unikn.publication.listelement.citation.prefix.version.undefined

Biochemistry. 1993, 32(51), pp. 14183-14186. ISSN 0006-2960. eISSN 1520-4995. Available under: doi: 10.1021/bi00214a017

Zusammenfassung

The bioluminescent reaction of luciferase isolated from Vibrio harveyi, strain M17, was initiated by mixing the luciferase-bound flavin 4a-hydroperoxide intermediate, purified in advance, with a long-chain aldehyde (dodecanal or octanal) at -4 °C . Measurements of absorbance changes from 300 to 600 nm during the course of the reaction revealed the existence of three sets of isosbestic points and three kinetic phases, the second of which parallels kinetically the decay of bioluminescence, measured concurrently. The absorbance changes in this second step and the decay of light emission exhibited similar deuterium isotope effects; this is postulated to be the step giving rise to the excited state and the enzyme-bound flavin 4a-hydroxide. The first step of the reaction, however, did not show an isotope effect; the intermediate thereby formed, observed here for the first time, is postulated to correspond to the luciferase-bound flavin 4a-peroxyhemiacetal.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Zitieren

ISO 690

MACHEROUX, Peter, Sandro GHISLA, J. Woodland HASTINGS, 1993. Spectral detection of an intermediate preceding the excited state in the bacterial luciferase reaction. In: Biochemistry. 1993, 32(51), pp. 14183-14186. ISSN 0006-2960. eISSN 1520-4995. Available under: doi: 10.1021/bi00214a017

BibTex

@article{Macheroux1993Spect-7947,
  year={1993},
  doi={10.1021/bi00214a017},
  title={Spectral detection of an intermediate preceding the excited state in the bacterial luciferase reaction},
  number={51},
  volume={32},
  issn={0006-2960},
  journal={Biochemistry},
  pages={14183--14186},
  author={Macheroux, Peter and Ghisla, Sandro and Hastings, J. Woodland}
}

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