AMP/GMP Analogs as Affinity ESIPT Probes for Highly Selective Sensing of Alkaline Phosphatase Activity in Living Systems

Chem Asian J. 2015 Nov;10(11):2444-51. doi: 10.1002/asia.201500280. Epub 2015 Aug 7.

Abstract

Current probes for alkaline phosphatase (ALP) detection had been developed mainly by adding a phosphate group to a dye, which would lead to indistinct performance when implemented in a living system as several phosphatases exist together. In this study, the nucleotides adenosine monophosphate (AMP) and guanosine monophosphate (GMP) were introduced into 2'-(2'-hydroxyphenyl)-benzothiazole-based probes, and highly fluorescent turn-on probes with good selectivity towards ALP over several phosphatases, as well as high affinity and low toxicity were obtained. In the presence of L-phenylalanine, an ALP inhibitor, a strong decrease in fluorescence recovery was observed. These probes allowed for real-time imaging of endogenous ALP activity in living cells as well as in a zebrafish model.

Keywords: AMP; GMP; alkaline phophatase; fluorescence; selectivity.

MeSH terms

  • Adenosine Monophosphate / analogs & derivatives*
  • Adenosine Monophosphate / metabolism
  • Alkaline Phosphatase / analysis
  • Alkaline Phosphatase / metabolism*
  • Animals
  • Benzothiazoles / chemistry
  • Benzothiazoles / metabolism
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • Guanosine Monophosphate / analogs & derivatives*
  • Guanosine Monophosphate / metabolism
  • HeLa Cells
  • Humans
  • Microscopy, Confocal
  • Zebrafish

Substances

  • Benzothiazoles
  • Fluorescent Dyes
  • Adenosine Monophosphate
  • Guanosine Monophosphate
  • Alkaline Phosphatase
  • benzothiazole