A water-soluble boronate-based fluorescent probe for the selective detection of peroxynitrite and imaging in living cells

Xiaolong Sun,Tony D James,John S Fossey,Juyoung Yoon,Steven D Bull,Gyoungmi Kim,John P Lowe,Qingling Xu,Stephen E Flower,Xuhong Qian

doi:10.1039/c4sc01417k

Abstract

A water-soluble boronate-based fluorescent probe was evaluated for the detection of peroxynitrite (ONOO−) in the presence of a monosaccharide. The enhanced fluorescence of the probe when bound with D-fructose was switched off in the presence of peroxynitrite. In contrast, other reactive oxygen/nitrogen species (ROS/RNS) led to only slight fluorescence decreases due to protection by an internal N–B interaction. The interaction of the probe with D-fructose not only strengthens the fluorescence signal, but also protects the boronic acid from oxidation by other ROS/RNS. Therefore, under conditions generating various ROS/RNS, the boronate-based saccharide complex preferentially reacts with peroxynitrite. The probe was used in cell imaging experiments for the detection of endogenous and exogenous peroxynitrite. The sensor displays good “on–off” responses towards peroxynitrite, both in RAW 264.7 cells and HeLa cells.

Highlights

Peroxynitrite (ONOOÀ) – a combination of nitric oxide and the superoxide radical anion – has been recognized as a strong oxidant in physiological and pathological processes
We investigated the selectivity of the 1–D-fructose complex towards other reactive oxygen and nitrogen species, such as hypochlorite (ÀOCl), nitric oxide (NO), nitrite (NO2À), nitrate (NO3À), peroxyl radicals (ROOc), superoxide (O2À) and hydroxyl radicals in pH 7.30 buffer solution (Fig. 4 and S8†)
We evaluated the ability of the probe 1–D-fructose complex to visualize exogenous and endogenous ONOOÀ using cell imaging experiments

Summary

Introduction

Peroxynitrite (ONOOÀ) – a combination of nitric oxide and the superoxide radical anion – has been recognized as a strong oxidant in physiological and pathological processes. The importance of peroxynitrite has led to researchers seeking effective and applicable approaches for its detection

Methods

Results

Conclusion