DNA Modifications

Norwood, C.B.; Vouros, Paul

doi:10.1007/978-1-4899-1748-5_3

Cited by 5 publications

(5 citation statements)

References 94 publications

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“…The exponent N may be as large as 1 for certain in vitro reactions but N is typically >6 for adducts formed in vivo. It has been estimated that approximately 500 mg of tissue is required for detection of DNA adducts if formed at levels of 1 in 10 6 and if the sensitivity of the detection is 100 pg (MW of adduct ≈ 500) …”

Section: Resultsmentioning

confidence: 99%

“…In recent years there has been an increased interest in the use of mass spectrometry for the analysis of DNA adducts. Mass spectrometry typically can provide structural information in the nanogram to picogram range and selective (nonstructural) detection in the picogram to femtogram range. − Therefore, given these characteristics, mass spectrometry is well positioned to play a significant role in the detection and characterization of DNA adducts. Our laboratory and others have pioneered the development of methodology based on the coupling of capillary separation techniques, such as liquid chromatography (LC) and CZE, with mass spectrometry for the analysis of DNA adducts. − CZE is a fast, low cost, and easy to use separation technique which also provides very high separation efficiencies.…”

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confidence: 99%

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Detection and Identification of Benzo[a]pyrene Diol Epoxide Adducts to DNA Utilizing Capillary Electrophoresis-Electrospray Mass Spectrometry

1996

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Capillary zone electrophoresis (CZE) coupled with negative ion electrospray mass spectrometry (ES-MS) is used for the detection and identification of adducts formed from the reaction of DNA with (+/-)-anti-7,8,9,10-tetrahydrobenzo[a]pyrene-7,8-diol 9,10-epoxide (BPDE),an active metabolite of benzo[a]pyrene (BaP). Results presented in this paper demonstrate low nanogram detection limits ( < 10 ng or < 15 pmol) for normal scan spectra and collision-induced dissociation spectra of the main nucleotide adduct formed from this reaction. (BPDE reacts predominantly with the exocyclic amino group of guanine.) Exploitation of selective reaction monitoring (SRM) produces detection limits in the low picogram range ( < 85 pg or < 130 fmol). The application of sample stacking significantly increases the concentration detection limit (to approximately 10(-8) M). Nucleotide adducts are negatively charged at most pHs and are therefore ideally suited to the stacking process used in this research. These techniques have been applied to the analysis of the adducts formed from the in vitro reaction of BPDE with DNA. In addition it is shown that CZE-ES-MS, combined with solid-phase sample cleanup, can detect adducts at levels of four adducts in 10(7) unmodified bases or less.

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Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

Detection and Identification of Benzo[a]pyrene Diol Epoxide Adducts to DNA Utilizing Capillary Electrophoresis-Electrospray Mass Spectrometry

1996

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“…Similarly, Giese et al () emphasized their own work in a short review on the same general subject but with more attention to method development and instrumental aspects of EC-MS. Detection of DNA adducts by all types of mass spectrometry has been reviewed recently ( − ). Similarly, another recent review has covered the detection of both DNA and protein adducts by general mass spectrometry ().…”

Section: Related Reviewsmentioning

confidence: 99%

Detection of DNA Adducts by Electron Capture Mass Spectrometry

Giese

1997

Chem. Res. Toxicol.

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“…NMR spectroscopy and X-ray crystallography enable detailed structural characterization but are time consuming and require large amounts (micromoles) of material. Mass spectrometry should complement these techniques by providing a rapid and sensitive means of determining the sequence selectivity of alkylation and, at the same time, provide some structural detail about the adducts .…”

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High Sensitivity and Fragmentation Specificity in the Analysis of Drug−DNA Adducts by Electrospray Tandem Mass Spectrometry

1997

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DNA alkylation is a key cellular event in the mechanism of action of clinical anticancer drugs, chemical mutagens, and carcinogens. 1 The nitrogen and oxygen atoms of purines are preferentially alkylated, with the affinity for a particular base depending on the sequence of neighboring bases. It is most likely that the sequence selectivity of the agent is important in determining the nature of the biological response. 2 Methods based on molecular biology have been used extensively to determine the sequence selectivity of alkylation. 3 Such assays are extremely sensitive and may be quantitative; however, they do not yield direct information concerning the structure of the ligand-DNA adducts. NMR spectroscopy 4 and X-ray crystallography 5 enable detailed structural characterization but are time consuming and require large amounts (micromoles) of material. Mass spectrometry should complement these techniques by providing a rapid and sensitive means of determining the sequence selectivity of alkylation and, at the same time, provide some structural detail about the adducts. 6 Several early studies concerned with the sequencing of ligand-DNA adducts by mass spectrometry have involved tandem mass spectrometry (MS/ MS) following fast atom bombardment (FAB) ionization. 7 This approach is limited, however, by the relatively poor efficiency of FAB ionization for oligonucleotides. 8 In contrast, electrospray ionization (ESI) has allowed mass measurement of relatively large oligonucleotides, including modified oligonucleotides, 9 an intact plasmid DNA, 10 and noncovalent drug-DNA complexes. 11 Tandem mass spectrometry following electrospray ionization (ESI-MS/MS) has been used for the location of modified bases within oligonucleotides and for the characterization of synthetic oligonucleotides which contain "unnatural" bases in their sequence. 12 To date, however, there have been no reports of the use of ESI-MS/MS for the sequencing of covalent, ligand-oligonucleotide adducts. 13 Hedamycin (Figure 1) is a naturally-occurring antitumor antibiotic that binds to double-stranded DNA by both intercalation and alkylation. DNA sequencing shows that alkylation occurs exclusively at guanine bases, but with a distinct preference for guanines located in 5′-CGT-3′ and 5′-CGG-3′ sequences. 14 NMR-derived solution structures of hedamycin covalently bound to double-stranded oligonucleotides show that the anthrapyrantrione chromophore of the ligand threads the double helix, placing the two amino sugars in the minor groove and the alkylating bis-epoxide side chain in the major groove. 15 Our previous studies of hedamycin-DNA binding by ESI-MS have confirmed that hedamycin significantly increases duplex stability and provided strong evidence for the formation of specifically, base-paired duplexes in the gas phase. 16 We demonstrate here that ESI-MS/MS can be used to determine unequivocally the sequence selectivity of base alkylation by a DNA-binding antitumor drug, using hedamycin as a model.ESI-MS/MS spectra of multiply-charged anions of u...

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DNA Modifications

Cited by 5 publications

References 94 publications

Detection and Identification of Benzo[a]pyrene Diol Epoxide Adducts to DNA Utilizing Capillary Electrophoresis-Electrospray Mass Spectrometry

Detection and Identification of Benzo[a]pyrene Diol Epoxide Adducts to DNA Utilizing Capillary Electrophoresis-Electrospray Mass Spectrometry

Detection of DNA Adducts by Electron Capture Mass Spectrometry

High Sensitivity and Fragmentation Specificity in the Analysis of Drug−DNA Adducts by Electrospray Tandem Mass Spectrometry

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