Inhibition of firefly luciferase by alkane analogues

Kô Takehara, Hiroshi Kamaya, Issaku Ueda

研究成果: Contribution to journalArticle査読

13 被引用数 (Scopus)

抄録

We reported that anesthetics increased the partial molal volume of firefly luciferase (FFL), while long-chain fatty acids (LCFA) decreased it. The present study measured the actions of dodecanol (neutral), dodecanoic acid (negatively charged), and dodecylamine (positively charged) hydrophobic molecules on FFL. The interaction modes are measured by (1) ATP-induced bioluminescence of FFL and (2) fluorescence of 2-(p-toluidino)naphthalene-6-sulfonate (TNS). TNS fluoresces brightly in hydrophobic media. It competes with the substrate luciferin on the FFL binding. From the Scatchard plot of TNS titration, the maximum binding number of TNS was 0.83, and its binding constant was 8.27×10 5 M -1. Job's plot also showed that the binding number is 0.89. From the TNS titration of FFL, the binding constant was estimated to be 8.8×10 5 M -1. Dodecanoic acid quenched the TNS fluorescence entirely. Dodecanol quenched about 25% of the fluorescence, whereas dodecylamine increased it. By comparing the fluorescence of TNS and bioluminescence of FFL, the binding modes and the inhibition mechanisms of these dodecane analogues are classified in three different modes: competitive (dodecanoic acid), noncompetitive (dodecylamine), and mixed (dodecanol).

本文言語英語
ページ(範囲)124-129
ページ数6
ジャーナルBiochimica et Biophysica Acta - General Subjects
1721
1-3
DOI
出版ステータス出版済み - 1 19 2005

All Science Journal Classification (ASJC) codes

  • 生物理学
  • 生化学
  • 分子生物学

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