Molecular modulated cysteine-selective fluorescent probe

Hyo Sung Jung, Tuhin Pradhan, Ji Hye Han, Kyung Jun Heo, Joung Hae Lee, Chulhun Kang, Jong Seung Kim

Research output: Contribution to journalArticle

124 Citations (Scopus)

Abstract

We have synthesized a series of coumarins (1-3) that can emit fluorescence in a turn-on manner through a Michael-type reaction with thiol-containing compounds. The only difference among the coumarins is the position of a carboxyl group on its benzene ring moiety near the double-bond conjugated coumarin. Their selectivity for Cys, GSH, and Hcy as well as the associated fluorogenic mechanism were illustrated by fluorescence spectroscopy, DFT calculations, and kinetic studies. All isomers prefer Cys over GSH in the reaction from 48.6 (probe 3) to 111-fold (probe 1) as demonstrated in a second order kinetics. The high selectivity of probe 1 to Cys might be achieved since the ortho carboxyl group on its benzene ring prefers a less negatively charged nucleophile. During intracellular Cys detection using 1, a possible interference by a large amount of GSH in the HepG2 cells was evaluated. The cells were treated with l-buthionine sulfoximine (BSO), an inhibitor of γ-glutamylcysteine synthetase, providing an experimental condition where the cells could not synthesize GSH from Cys or other species. Then, the fluorescence intensity of 1 in HepG2 cells under BSO-H2O2 treatment was strongly enhanced by N-acetylcysteine (NAC), a precursor of Cys, implicating that the fluorescence signal from the cells is mainly associated with changes in intracellular [Cys] rather than that in intracellular [GSH].

Original languageEnglish
Pages (from-to)8495-8502
Number of pages8
JournalBiomaterials
Volume33
Issue number33
DOIs
Publication statusPublished - 2012 Nov 1

Keywords

  • Cellular detection
  • Cysteine
  • DFT calculations
  • Fluorescence
  • Thiol

ASJC Scopus subject areas

  • Biomaterials
  • Bioengineering
  • Ceramics and Composites
  • Mechanics of Materials
  • Biophysics

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  • Cite this

    Jung, H. S., Pradhan, T., Han, J. H., Heo, K. J., Lee, J. H., Kang, C., & Kim, J. S. (2012). Molecular modulated cysteine-selective fluorescent probe. Biomaterials, 33(33), 8495-8502. https://doi.org/10.1016/j.biomaterials.2012.08.009