Development of luminescent coelenterazine derivatives activatable by β-galactosidase for monitoring dual gene expression

Eric Lindberg, Shin Mizukami, Keiji Ibata, Atsushi Miyawaki, Kazuya Kikuchi

研究成果: Article査読

20 被引用数 (Scopus)

抄録

Two bioluminogenic caged coelenterazine derivatives (bGalCoel and bGalNoCoel) were designed and synthesized to detect β-galactosidase activity and expression by means of bioluminescence imaging. Our approach addresses the instability of coelenterazine by introducing β-galactose caging groups to block the auto-oxidation of coelenterazine. Both probes contain β-galactosidase cleavable caging groups at the carbonyl group of the imidazo-pyrazinone moiety. One of the probes in particular, bGalNoCoel, displayed a fast cleavage profile, high stability, and high specificity for β-galactosidase over other glycoside hydrolases. bGalN-oCoel could detect β-galactosidase activity in living HEK-293T cell cultures that expressed a mutant Gaussia luciferase. It was determined that coelenterazine readily diffuses in and out of cells after uncaging by β-galactosidase. We showed that this new caged coelenterazine derivative, bGalNoCoel, could function as a dual-enzyme substrate and detect enzyme activity across two separate cell populations.

本文言語English
ページ(範囲)14970-14976
ページ数7
ジャーナルChemistry - A European Journal
19
44
DOI
出版ステータスPublished - 2013 10 25
外部発表はい

ASJC Scopus subject areas

  • Catalysis
  • Organic Chemistry

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