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DNA Adductomics by mass tag prelabeling.

Poguang Wang1, Elisabeth Roider2, Michael E Coulter3

  • 1Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, USA.

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|April 6, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces a novel method for DNA adductomics, enabling the detection of polar DNA adducts using cationic xylyl (CAX) mass tags. This technique offers a practical approach to analyzing DNA damage and repair mechanisms.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Analytical Chemistry

Background:

  • DNA adductomics is crucial for understanding DNA damage and repair.
  • Existing methods face challenges in detecting polar DNA adducts.
  • Nucleotide excision repair mechanisms recognize DNA structural disruptions.

Purpose of the Study:

  • To develop a new approach for DNA adductomics using direct alkylation of double-stranded DNA.
  • To enable the practical detection of polar DNA adducts.
  • To utilize the DNA structural disruption caused by adducts for preferential labeling.

Main Methods:

  • Intact double-stranded DNA was reacted with a cationic xylyl (CAX) mass tag at 45°C.
  • Adducted nucleotides were detected using liquid chromatography/mass spectrometry (LC/MS²).
  • Enzymatic digestion and solid-phase extraction were employed for sample preparation.

Main Results:

  • A diversity of 42 DNA or RNA adducts were detected, including 8-oxo-dGMP and benzoquinone-hydroxymethyl-dCMP.
  • The method successfully detected polar adducts in a single procedure.
  • Preliminary sensitivity analysis indicated detection of an adduct at approximately 4 adducts per 10^9 nucleotides.

Conclusions:

  • CAX-Prelabeling is a novel and practical technique for polar DNA adductomics.
  • The method offers a new avenue for studying DNA damage and repair.
  • Further research is needed to optimize scope and sensitivity.