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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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Application of AlDeSense to Stratify Ovarian Cancer Cells Based on Aldehyde Dehydrogenase 1A1 Activity
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Detecting ALDH2 activity in live cells via conditional metabolic labeling.

Weisong Lv1,2, Zheng Wang1,2, Can Zhang2

  • 1College of Pharmaceutical Sciences, Zhejiang University 866 Yuhangtang Street Hangzhou 310058 China fanxh@zju.edu.cn lixin81@zju.edu.cn.

Chemical Science
|August 25, 2025
PubMed
Summary

We developed a new method called conditional metabolic labeling for enzymatic activity detection (cMLEAD) to measure enzyme function in living cells. This strategy reliably detects aldehyde dehydrogenase 2 (ALDH2) activity and aids in discovering new drugs.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Chemical Biology

Background:

  • Detecting enzyme activity in live cells is challenging.
  • Existing metabolic labeling methods lack specificity.
  • Enzyme activity is crucial for cellular function and disease.

Purpose of the Study:

  • To introduce a novel strategy, cMLEAD, for detecting specific enzyme activity in live cells.
  • To demonstrate the utility of cMLEAD for measuring aldehyde dehydrogenase 2 (ALDH2) activity.
  • To establish cMLEAD as a platform for drug discovery and understanding enzyme function in disease.

Main Methods:

  • Developed cMLEAD using an azido-tagged precursor metabolized by ALDH2.
  • Utilized click chemistry for fluorescence-based detection of metabolic incorporation.
  • Validated ALDH2 activity measurements through genetic and pharmacological modulation.
  • Applied cMLEAD to study ALDH2 activity in cellular senescence and oxidative stress.

Main Results:

  • cMLEAD reliably detects ALDH2 activity in live cells.
  • ALDH2 activity was suppressed in cellular senescence and oxidative stress.
  • Identified sennoside A as an ALDH2 activator using a cMLEAD-based screening platform.
  • Sennoside A alleviated light-induced retinal degeneration in mice.

Conclusions:

  • cMLEAD is a robust and versatile platform for enzyme activity detection in native cellular contexts.
  • cMLEAD facilitates the discovery of enzyme modulators with therapeutic potential.
  • The cMLEAD strategy can be adapted for detecting other enzyme activities.