Intracellular Fate of 2-NBDG, a Fluorescent Probe for Glucose Uptake Activity, in Escherichia Coli Cells

Biosci Biotechnol Biochem. 1996 Nov;60(11):1899-901. doi: 10.1271/bbb.60.1899.

Abstract

A fluorescent derivative of D-glucose, 2-NBDG, which was previously developed for the evaluation of glucose uptake activity by living cells, was used on Escherichia coli cells and its fate after incorporation in the cells was investigated. 2-NBDG was converted to another fluorescent derivative (2-NBDG metabolite) immediately after it was taken by E. coli cells. This 2-NBDG metabolite was then decomposed to non-fluorescent forms. 2-NBDG metabolite was decomposed into the original 2-NBDG by G6Pase with concurrent liberation of inorganic phosphate. Furthermore, FAB/MS analysis showed that its molecular weight was 420, the same value as that of 2-NBDG 6-phosphate. These indicate 2-NBDG metabolite should be 2-NBDG 6-phosphate. Based on these results, the feasibility of 2-NBDG as a fluorescent non-toxic probe for glucose uptake activity and its application to viability assessment of various living systems are discussed.

MeSH terms

  • 4-Chloro-7-nitrobenzofurazan / analogs & derivatives*
  • 4-Chloro-7-nitrobenzofurazan / metabolism
  • Chromatography, High Pressure Liquid
  • Deoxyglucose / analogs & derivatives*
  • Deoxyglucose / metabolism
  • Escherichia coli / chemistry
  • Escherichia coli / metabolism*
  • Fluorescent Dyes / metabolism*
  • Glucose / metabolism*
  • Spectrometry, Fluorescence
  • Spectrometry, Mass, Fast Atom Bombardment

Substances

  • Fluorescent Dyes
  • Deoxyglucose
  • 4-Chloro-7-nitrobenzofurazan
  • Glucose
  • 2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose