Combined high-performance liquid chromatography/ sup 32 P-postlabeling assay of N7-methyldeoxyguanosine
Journal Article
·
· Cancer Research; (USA)
OSTI ID:6284167
- National Cancer Institute, NIH, Bethesda, MD (USA)
A highly sensitive and specific assay for the detection of N7-methyl-2{prime}-deoxyguanosine (N7methyldG) has been developed by combining high-performance liquid chromatography, {sup 32}P-postlabeling, and nucleotide chromatography. Separation of normal nucleotides and adducts by high-performance liquid chromatography and then combining a portion of 2{prime}-deoxyguanosine to the N7methyldG allows for quantitation using an internal standard. The directly determined molar ratio is not subject to errors in digestion, variable ATP-specific activity, or assumptions in relative adduct-labeling efficiency. The detection limit was one N7methyldG adduct in 10(7) unmodified 2{prime}-deoxyguanosine bases. N7methyldG adducts have been detected in 5 human lung samples in which O6-methyl-2{prime}-deoxyguanosine adducts had been previously determined. The mean ratio of N7methyldG to O6-methyl-2{prime}-deoxyguanosine was determined to be approximately 10. The current assay complements the high-performance liquid chromatography/{sup 32}P-postlabeling assay for O6-methyl-2{prime}-deoxyguanosine and increases the detection sensitivity of DNA methylated by exogenous alkylating agents.
- OSTI ID:
- 6284167
- Journal Information:
- Cancer Research; (USA), Journal Name: Cancer Research; (USA) Vol. 50:20; ISSN 0008-5472; ISSN CNREA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADDUCTS
ALKYLATING AGENTS
ANIMALS
AROMATICS
ATP
AZAARENES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DNA ADDUCTS
GUANOSINE
HETEROCYCLIC COMPOUNDS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIQUID COLUMN CHROMATOGRAPHY
LUNGS
MAMMALS
MAN
METHYLATION
NUCLEI
NUCLEOSIDES
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PRIMATES
PURINES
RADIOASSAY
RADIOISOTOPES
RESPIRATORY SYSTEM
RIBOSIDES
SEPARATION PROCESSES
TRACER TECHNIQUES
VERTEBRATES
59 BASIC BIOLOGICAL SCIENCES
ADDUCTS
ALKYLATING AGENTS
ANIMALS
AROMATICS
ATP
AZAARENES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BODY
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DNA ADDUCTS
GUANOSINE
HETEROCYCLIC COMPOUNDS
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
LIQUID COLUMN CHROMATOGRAPHY
LUNGS
MAMMALS
MAN
METHYLATION
NUCLEI
NUCLEOSIDES
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PRIMATES
PURINES
RADIOASSAY
RADIOISOTOPES
RESPIRATORY SYSTEM
RIBOSIDES
SEPARATION PROCESSES
TRACER TECHNIQUES
VERTEBRATES