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

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

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)14970-14976
Number of pages7
JournalChemistry - A European Journal
Volume19
Issue number44
DOIs
Publication statusPublished - 2013 Oct 25

Keywords

  • biosensors
  • cleavage reactions
  • enzymes
  • galactosidase
  • luminescence

ASJC Scopus subject areas

  • Catalysis
  • Organic Chemistry

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