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

16 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 (Weinheim an der Bergstrasse, Germany)
Volume19
Issue number44
DOIs
Publication statusPublished - 2013 Oct 25
Externally publishedYes

Fingerprint

Galactosidases
Gene Expression Profiling
Gene expression
Derivatives
Monitoring
Bioluminescence
HEK293 Cells
Glycoside Hydrolases
Enzyme activity
Enzymes
Luciferases
Galactose
Cell culture
Cell Culture Techniques
Cells
coelenterazine
Imaging techniques
Oxidation
Substrates
Population

Keywords

  • biosensors
  • cleavage reactions
  • enzymes
  • galactosidase
  • luminescence

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Development of luminescent coelenterazine derivatives activatable by β-galactosidase for monitoring dual gene expression. / Lindberg, Eric; Mizukami, Shin; Ibata, Keiji; Miyawaki, Atsushi; Kikuchi, Kazuya.

In: Chemistry (Weinheim an der Bergstrasse, Germany), Vol. 19, No. 44, 25.10.2013, p. 14970-14976.

Research output: Contribution to journalArticle

Lindberg, Eric ; Mizukami, Shin ; Ibata, Keiji ; Miyawaki, Atsushi ; Kikuchi, Kazuya. / Development of luminescent coelenterazine derivatives activatable by β-galactosidase for monitoring dual gene expression. In: Chemistry (Weinheim an der Bergstrasse, Germany). 2013 ; Vol. 19, No. 44. pp. 14970-14976.
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