Synthesis and 32P-postlabeling/high-performance liquid chromatography separation of diastereomeric 1,N2-(1,3-propano)-2'-deoxyguanosine 3'-phosphate adducts formed from 4-hydroxy-2-nonenal.
4-Hydroxy-2-nonenal (HNE), a major electrophilic byproduct of lipid peroxidation, is mutagenic and cytotoxic. The two pairs of HNE-derived diastereomeric 1,N2-propanodeoxyguanosine 3'-monophosphate adducts were synthesized from reaction of HNE with 2'-deoxyguanosine 3'-monophosphate. After HPLC separation, these adducts were characterized by UV-visible absorption and negative ion electrospray ionization MS/MS analysis. To further characterize the structures, these adducts were dephosphorylated to the corresponding HNE-modified deoxyguanosine adducts and their HPLC retention times and UV spectra were compared with those of the synthetic standards prepared from reaction of HNE with 2'-deoxyguanosine. Separation of these adducts by 32P-postlabeling/HPLC was developed. Reaction of HNE with calf thymus DNA resulted in only one pair of diastereomeric adducts, with one adduct predominantly formed with a modification level of 1.2 +/- 0.5 adducts/10(7) nucleotides.
Yi
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P
National Center for Toxicological Research, Jefferson, Arkansas 72079, USA.
Zhan
D
D
Samokyszyn
V M
VM
Doerge
D R
DR
Fu
P P
PP
D000447:Aldehydes / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D015853:Cysteine Proteinase Inhibitors / Q000737:chemistry*; D018736:DNA Adducts / Q000737:chemistry*; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry / Q000302:isolation & purification; D007553:Isotope Labeling; D008279:Magnetic Resonance Imaging; D008968:Molecular Conformation; D009153:Mutagens / Q000138:chemical synthesis* / Q000302:isolation & purification*; D010761:Phosphorus Radioisotopes; D013056:Spectrophotometry, Ultraviolet; D013237:Stereoisomerism
C090855:1,N(2)-(1,3-propano)-2'-deoxyguanosine; D000447:Aldehydes; D015853:Cysteine Proteinase Inhibitors; D018736:DNA Adducts; D009153:Mutagens; D010761:Phosphorus Radioisotopes; D003849:Deoxyguanosine; C027576:4-hydroxy-2-nonenal