Design and synthesis of highly sensitive and selective fluorescein-derived magnesium fluorescent probes and application to intracellular 3D Mg2+ imaging

Hirokazu Komatsu, Naoko Iwasawa, Daniel Citterio, Yoshio Suzuki, Takeshi Kubota, Kentaro Tokuno, Yoshiichiro Kitamura, Kotaro Oka, Koji Suzuki

Research output: Contribution to journalArticle

125 Citations (Scopus)

Abstract

The role of intracellular magnesium ions is of high interest in the fields of pharmacology and cellular biology. To accomplish the dynamic and three-dimensional imaging of intracellular Mg2+, there is a strong desire for the development of optimized Mg2+ fluorescent probes. In this paper we describe the design, synthesis, and cellular application of the three novel Mg2+ fluorescent probes KMG-101, -103, and -104. The compounds of this series feature a charged β-diketone as a binding site specific for Mg2+ and a fluorescein residue as the fluorophore that can be excited with an Ar+ laser such as is widely used in confocal scanning microscopy. This molecular design leads to an intensive off-on-type fluorescent response toward Mg2+ ions. The two fluorescent probes KMG-103 and -104 showed suitable dissociation constants (Kd,Mg2+ = 2 mM) and nearly a 10-fold fluorescence enhancement over the intracellular magnesium ion concentration range (0.1-6 mM), allowing high-contrast, sensitive, and selective Mg2+ measurements. For intracellular applications, the membrane-permeable probe KMG-104AM was synthesized and successfully incorporated into PC12 cells. Upon application of the mitochondria uncoupler FCCP to the probe-incorporated cells, the resulting increase in the free magnesium ion concentration could be followed over time. By using a confocal microscope, the intracellular 3D magnesium ion concentration distributions were satisfactorily observed.

Original languageEnglish
Pages (from-to)16353-16360
Number of pages8
JournalJournal of the American Chemical Society
Volume126
Issue number50
DOIs
Publication statusPublished - 2004 Dec 22

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Fluorescein
Fluorescent Dyes
Magnesium
Ions
Imaging techniques
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Cytology
Intracellular Membranes
Mitochondria
Three-Dimensional Imaging
Fluorophores
PC12 Cells
Binding sites
Confocal Microscopy
Cell Biology
Microscopic examination
Lasers
Microscopes
Fluorescence
Binding Sites

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Design and synthesis of highly sensitive and selective fluorescein-derived magnesium fluorescent probes and application to intracellular 3D Mg2+ imaging. / Komatsu, Hirokazu; Iwasawa, Naoko; Citterio, Daniel; Suzuki, Yoshio; Kubota, Takeshi; Tokuno, Kentaro; Kitamura, Yoshiichiro; Oka, Kotaro; Suzuki, Koji.

In: Journal of the American Chemical Society, Vol. 126, No. 50, 22.12.2004, p. 16353-16360.

Research output: Contribution to journalArticle

Komatsu, Hirokazu ; Iwasawa, Naoko ; Citterio, Daniel ; Suzuki, Yoshio ; Kubota, Takeshi ; Tokuno, Kentaro ; Kitamura, Yoshiichiro ; Oka, Kotaro ; Suzuki, Koji. / Design and synthesis of highly sensitive and selective fluorescein-derived magnesium fluorescent probes and application to intracellular 3D Mg2+ imaging. In: Journal of the American Chemical Society. 2004 ; Vol. 126, No. 50. pp. 16353-16360.
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