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Detection of biothiols in cells by a terbium chelate-Hg (II) system.

Abstract
Great efforts have been devoted to the development of sensitive and specific analysis methods for biothiols because of their important roles in biological systems. We present a new detection system for biothiols that is based on the reversible quenching and restoration of fluorescence of terbium chelate caused by Hg(2+) and thiol species. In the presence of biothiols, a restoration of fluorescence of terbium chelate after quenching by Hg(2+) was observed due to the interaction of Hg(2+) with thiol groups, and the restored fluorescence increased with the concentration of biothiols. This method was sensitive and selective for biothiols. The detection limit was 80 nM for glutathione, 100 nM for Hcy, and 400 nM for Cysteine, respectively. The terbium chelate-Hg (II) system was successfully applied to determine the levels of biothiols in cancer cells and urine samples. Further, it was also shown to be comparable to Ellman's assay. Compared to other fluorescence methods, the terbium chelate probe is advantageous because interference from short-lived nonspecific fluorescence can be efficiently eliminated due to the long fluorescence lifetime of terbium chelate, which allows for detection by time-resolved fluorescence. The terbium chelate probe can serve as a diagnostic tool for the detection of abnormal levels of biothiols in disease.
AuthorsHongliang Tan, Yang Chen
JournalJournal of biomedical optics (J Biomed Opt) Vol. 17 Issue 1 Pg. 017001 (Jan 2012) ISSN: 1560-2281 [Electronic] United States
PMID22352667 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Coordination Complexes
  • Mercury Compounds
  • Molecular Probes
  • Sulfhydryl Compounds
  • Terbium
  • Homocysteine
  • Glutathione
  • Cysteine
Topics
  • Cell Line, Tumor
  • Coordination Complexes (chemistry)
  • Cysteine (analysis, chemistry)
  • Glutathione (analysis, chemistry)
  • Histocytochemistry
  • Homocysteine (analysis, chemistry)
  • Humans
  • Mercury Compounds (chemistry)
  • Molecular Probes (chemistry)
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence (methods)
  • Sulfhydryl Compounds (analysis, chemistry)
  • Terbium (chemistry)
  • Urine (chemistry)

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