Design and synthesis of Mg2+-selective fluoroionophores based on a coumarin derivative and application for Mg2+ measurement in a living cell

Yoshio Suzuki, Hirokazu Komatsu, Takafumi Ikeda, Naohiko Saito, Sawa Araki, Daniel Citterio, Hideaki Hisamoto, Yoshiichiro Kitamura, Takeshi Kubota, Jun Nakagawa, Kotaro Oka, Koji Suzuki

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Abstract

Novel Mg2+ fluorescent molecular probes (KMG-20-AM and KMG-27-AM; where AM is an acetoxymethyl group) based on a coumarin possessing a charged β-diketone structure were designed and synthesized. These fluorescent probes produced a red shift from 425 to 445 nm in the absorption spectra after formation of a complex with Mg2+. The fluorescence spectra of these probes also showed a red shift from 485 to 495 nm and an increasing fluorescence intensity after formation of a complex with Mg2+. The optimum experimental conditions were excitation wavelength of 445 nm and a monitored wavelength of 500 nm, where these probes functioned as an indicator showing an image of increasing fluorescence in the presence of Mg2+. These probes showed a "seesaw-type" fluorescent spectral change with the isosbestic point at 480 nm due to the light excitation at 445 nm, which indicates that ratiometry can be used for the measurement. The molecular probes formed a 1:1 complex with Mg2+ and the dissociation constant (Kd) was 10.0 mM for KMG-20. The association constants of the probes with Mg2+ were ∼3 times higher than that with Ca2+, which showed that the selectivity of Mg2+ versus Ca2+ for these probes was over 200 times higher than that for commercially available Mg2+ fluorescent molecular probes such as mag-fura-2, Magnesium Green. As an application of these probes, intracellular fluorescent imaging of Mg2+ was demonstrated using a fluorescent microscope. After the addition of KMG-20-AM and KMG-27-AM into PC12 cells, a strong fluorescence was observed in the cytoplasm and a weak fluorescence in the nuclei region. After treatment with a high-K+ medium, the fluorescence intensity increased due to increasing intracellular Mg2+. The real image of Mg2+ release from the magnesium store was successfully observed with these Mg2+ fluorescent probes.

Original languageEnglish
Pages (from-to)1423-1428
Number of pages6
JournalAnalytical Chemistry
Volume74
Issue number6
DOIs
Publication statusPublished - 2002 Mar 15

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Coumarins
Fluorescence
Cells
Molecular Probes
Fluorescent Dyes
Wavelength
Magnesium
Absorption spectra
Microscopes
Association reactions
Imaging techniques

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Design and synthesis of Mg2+-selective fluoroionophores based on a coumarin derivative and application for Mg2+ measurement in a living cell. / Suzuki, Yoshio; Komatsu, Hirokazu; Ikeda, Takafumi; Saito, Naohiko; Araki, Sawa; Citterio, Daniel; Hisamoto, Hideaki; Kitamura, Yoshiichiro; Kubota, Takeshi; Nakagawa, Jun; Oka, Kotaro; Suzuki, Koji.

In: Analytical Chemistry, Vol. 74, No. 6, 15.03.2002, p. 1423-1428.

Research output: Contribution to journalArticle

Suzuki, Y, Komatsu, H, Ikeda, T, Saito, N, Araki, S, Citterio, D, Hisamoto, H, Kitamura, Y, Kubota, T, Nakagawa, J, Oka, K & Suzuki, K 2002, 'Design and synthesis of Mg2+-selective fluoroionophores based on a coumarin derivative and application for Mg2+ measurement in a living cell', Analytical Chemistry, vol. 74, no. 6, pp. 1423-1428. https://doi.org/10.1021/ac010914j
Suzuki, Yoshio ; Komatsu, Hirokazu ; Ikeda, Takafumi ; Saito, Naohiko ; Araki, Sawa ; Citterio, Daniel ; Hisamoto, Hideaki ; Kitamura, Yoshiichiro ; Kubota, Takeshi ; Nakagawa, Jun ; Oka, Kotaro ; Suzuki, Koji. / Design and synthesis of Mg2+-selective fluoroionophores based on a coumarin derivative and application for Mg2+ measurement in a living cell. In: Analytical Chemistry. 2002 ; Vol. 74, No. 6. pp. 1423-1428.
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AU - Saito, Naohiko

AU - Araki, Sawa

AU - Citterio, Daniel

AU - Hisamoto, Hideaki

AU - Kitamura, Yoshiichiro

AU - Kubota, Takeshi

AU - Nakagawa, Jun

AU - Oka, Kotaro

AU - Suzuki, Koji

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