Detection of NAD(P)H-dependent enzyme activity by time-domain ratiometry of terbium luminescence

Takuya Terai, Hiroki Ito, Kenjiro Hanaoka, Toru Komatsu, Tasuku Ueno, Tetsuo Nagano, Yasuteru Urano

研究成果: Article査読

9 被引用数 (Scopus)

抄録

NAD(P)H-dependent oxidoreductases play important roles in biology. Recently, we reported that the luminescence lifetime of some Tb3+ complexes is sensitive to NAD(P)H, and we used this phenomenon to detect activities of these enzymes. However, conventional time-resolved luminescence assays are susceptible to static quenchers such as ATP. Herein we describe a detection methodology that overcomes this issue: the intensity of the sample is measured twice with different delay times and the intensity ratio value is used as an index of NAD(P)H concentration. The method is more robust than single-point measurement, and is compatible with high-throughput assays using conventional microplate readers.

本文言語English
ページ(範囲)2314-2317
ページ数4
ジャーナルBioorganic and Medicinal Chemistry Letters
26
9
DOI
出版ステータスPublished - 2016 5月 1
外部発表はい

ASJC Scopus subject areas

  • 生化学
  • 分子医療
  • 分子生物学
  • 薬科学
  • 創薬
  • 臨床生化学
  • 有機化学

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