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DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
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Nitroxyl donating and visualization with a coumarin-based fluorescence probe.

Jiajun Chen1, Yunxi Cui2, Peixuan Wu1

  • 1Key Laboratory of Advanced Materials of Tropical Island Resources of Ministry of Education and School of Chemical Engineering and Technology, Hainan University, Haikou, Hainan 570228, China.

Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
|May 1, 2024
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel coumarin-based compound (CD1) that releases both nitroxyl (HNO) and a fluorescent signal. This dual-functional donor allows for real-time monitoring of HNO release in biological systems.

Keywords:
Cell imagingDonor moleculeFluorescenceNitroxyl (HNO)Probe

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Area of Science:

  • Chemistry
  • Biomedical Engineering
  • Pharmacology

Background:

  • Nitroxyl (HNO) is a signaling molecule with therapeutic potential for conditions like congestive heart failure.
  • Existing methods for detecting HNO release can be limited in real-time monitoring and biological application.
  • Development of novel HNO donors is crucial for advancing HNO-related research and therapeutic strategies.

Purpose of the Study:

  • To synthesize and characterize a novel coumarin-based compound (CD1) as a dual-functional HNO donor.
  • To investigate the HNO release kinetics and stoichiometry of CD1 under physiological conditions.
  • To demonstrate the utility of CD1 for in situ monitoring of HNO and intracellular bioimaging.

Main Methods:

  • Synthesis of N-hydroxy-2-oxo-2H-chromene-6-sulfonamide (CD1).
  • Decomposition studies of CD1 at pH 7.4 and 37°C to determine half-life.
  • Confirmation of HNO stoichiometry using Vitamin B12 and phosphine traps.
  • Fluorescence monitoring of the degradation product (CS1) for HNO quantification.
  • Cellular bioimaging in HeLa cells to visualize intracellular HNO release.

Main Results:

  • CD1 demonstrated a half-life of approximately 90 minutes under physiological conditions.
  • Stoichiometric release of HNO from CD1 was confirmed.
  • The degradation product, 2-oxo-2H-chromene-6-sulfinate (CS1), served as a reliable fluorescent marker for HNO.
  • A 4.9-fold increase in fluorescence intensity correlated with HNO release, with a detection limit of ~0.13 μM.
  • Intracellular fluorescence signals were observed in HeLa cells, indicating successful bioimaging of HNO release.

Conclusions:

  • The novel coumarin-based CD1 compound functions as an effective dual-functional HNO donor.
  • CD1 enables accurate, real-time monitoring of HNO release through fluorescence generation.
  • CD1 holds promise as a valuable tool for exploring HNO biology and potential therapeutic applications.