Measurement of oxidatively induced DNA damage and its repair, by mass spectrometric techniques
M Dizdaroglu1, E Coskun, P Jaruga
1Biomolecular Measurement Division, National Institute of Standards and Technology , Gaithersburg, MD , USA.
Abstract:
Oxidatively induced damage caused by free radicals and other DNA-damaging agents generate a plethora of products in the DNA of living organisms. There is mounting evidence for the involvement of this type of damage in the etiology of numerous diseases including carcinogenesis. For a thorough understanding of the mechanisms, cellular repair, and biological consequences of DNA damage, accurate measurement of resulting products must be achieved. There are various analytical techniques, with their own advantages and drawbacks, which can be used for this purpose. Mass spectrometric techniques with isotope dilution, which include gas chromatography (GC) and liquid chromatography (LC), provide structural elucidation of products and ascertain accurate quantification, which are absolutely necessary for reliable measurement. Both gas chromatography-mass spectrometry (GC-MS) or liquid chromatography-mass spectrometry (LC-MS), in single or tandem versions, have been used for the measurement of numerous DNA products such as sugar and base lesions, 8,5'-cyclopurine-2'-deoxynucleosides, base-base tandem lesions, and DNA-protein crosslinks, in vitro and in vivo. This article reviews these techniques and their applications in the measurement of oxidatively induced DNA damage and its repair.
Insights
Accurate measurement of DNA damage is crucial for understanding diseases like cancer. Mass spectrometric techniques, including GC-MS and LC-MS, are essential for quantifying these oxidative DNA products and their repair.
Area of Science:
- Biochemistry
- Molecular Biology
- Analytical Chemistry
Background:
- Oxidative stress generates DNA damage, implicated in diseases such as cancer.
- Understanding DNA damage mechanisms, repair, and consequences requires precise measurement of damage products.
- Various analytical techniques exist, each with limitations.
Purpose of the Study:
- To review analytical techniques for measuring oxidatively induced DNA damage.
- To highlight the role of mass spectrometry in quantifying DNA damage products.
- To discuss the application of these techniques in studying DNA repair.
Main Methods:
- Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) are discussed.
- Isotope dilution techniques are employed for accurate quantification.
- Single and tandem mass spectrometry versions are utilized.
Main Results:
- GC-MS and LC-MS enable structural elucidation and accurate quantification of DNA damage products.
- These methods have been applied to measure various lesions, including base and sugar modifications, cyclopurine deoxynucleosides, tandem lesions, and DNA-protein crosslinks.
- The techniques are effective for both in vitro and in vivo studies.
Conclusions:
- Mass spectrometric techniques with isotope dilution are vital for reliable measurement of oxidatively induced DNA damage.
- GC-MS and LC-MS are powerful tools for investigating DNA damage and repair mechanisms.
- Accurate quantification is essential for advancing our understanding of DNA damage's role in disease etiology.
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