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Natalia Y Tretyakova - One of the best experts on this subject based on the ideXlab platform.
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reversible dna Protein Cross Linking at epigenetic dna marks
Angewandte Chemie, 2017Co-Authors: Hongzhao Shao, Qiyuan Han, Christopher L Seiler, Natalia Y TretyakovaAbstract:5-Formylcytosine (5fC) is an endogenous DNA modification frequently found within regulatory elements of mammalian genes. Although 5fC is an oxidation product of 5-methylcytosine (5mC), the two epigenetic marks show distinct genome-wide distributions and Protein affinities, suggesting that they perform different functions in epigenetic signaling. A unique feature of 5fC is the presence of a potentially reactive aldehyde group in its structure. Here, we show that 5fC bases in DNA readily form Schiff base conjugates with Lys side chains of nuclear Proteins in vitro and in vivo. These covalent Protein-DNA complexes are reversible (t1/2, 1.8 h), suggesting that they contribute to transcriptional regulation and chromatin remodeling. On the other hand, 5fC mediated DNA-Protein Cross-links, if present at replication forks or actively transcribed regions, may interfere with DNA replication and transcription.
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mass spectrometry based proteomics study of cisplatin induced dna Protein Cross Linking in human fibrosarcoma ht1080 cells
Chemical Research in Toxicology, 2017Co-Authors: Xun Ming, Arnold Groehler, Erin D Michaelsonrichie, Peter W Villalta, Colin R Campbell, Natalia Y TretyakovaAbstract:Platinum-based antitumor drugs such as 1,1,2,2-cis-diamminedichloroplatinum(II) (cisplatin), carboplatin, and oxaliplatin are currently used to treat nearly 50% of all cancer cases, and novel platinum based agents are under development. The antitumor effects of cisplatin and other platinum compounds are attributed to their ability to induce interstrand DNA–DNA Cross-links, which are thought to inhibit tumor cell growth by blocking DNA replication and/or preventing transcription. However, platinum agents also induce significant numbers of unusually bulky and helix-distorting DNA–Protein Cross-links (DPCs), which are poorly characterized because of their unusual complexity. We and others have previously shown that DPCs block DNA replication and transcription and causes toxicity in human cells, potentially contributing to the biological effects of platinum agents. In the present work, we have undertaken a system-wide investigation of cisplatin-mediated DNA–Protein Cross-Linking in human fibrosarcoma (HT1080)...
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mass spectrometry based tools to characterize dna Protein Cross Linking by bis electrophiles
Basic & Clinical Pharmacology & Toxicology, 2017Co-Authors: Arnold Groehler, Amanda Degner, Natalia Y TretyakovaAbstract:DNA-Protein Cross-links (DPCs) are unusually bulky DNA adducts that form in cells as a result of exposure to endogenous and exogenous agents including reactive oxygen species, ultraviolet light, ionizing radiation, environmental agents (e.g. transition metals, formaldehyde, 1,2-dibromoethane, 1,3-butadiene) and common chemotherapeutic agents. Covalent DPCs are cytotoxic and mutagenic due to their ability to interfere with faithful DNA replication and to prevent accurate gene expression. Key to our understanding of the biological significance of DPC formation is identifying the Proteins most susceptible to forming these unusually bulky and complex lesions and quantifying the extent of DNA-Protein Cross-Linking in cells and tissues. Recent advances in bottom-up mass spectrometry-based proteomics have allowed for an unbiased assessment of the whole Protein DPC adductome after in vitro and in vivo exposures to Cross-Linking agents. This MiniReview summarizes current and emerging methods for DPC isolation and analysis by mass spectrometry-based proteomics. We also highlight several examples of successful applications of these novel methodologies to studies of DPC lesions induced by bis-electrophiles such as formaldehyde, 1,2,3,4-diepoxybutane, nitrogen mustards and cisplatin.
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covalent dna Protein Cross Linking by phosphoramide mustard and nornitrogen mustard in human cells
Chemical Research in Toxicology, 2016Co-Authors: Arnold Groehler, Peter W Villalta, Colin R Campbell, Natalia Y TretyakovaAbstract:N,N-Bis-(2-chloroethyl)-phosphorodiamidic acid (phosphoramide mustard, PM) and N,N-bis-(2-chloroethyl)-amine (nornitrogen mustard, NOR) are the two biologically active metabolites of cyclophosphamide, a DNA alkylating drug commonly used to treat lymphomas, breast cancer, certain brain cancers, and autoimmune diseases. PM and NOR are reactive bis-electrophiles capable of Cross-Linking cellular biomolecules to form covalent DNA–DNA and DNA–Protein Cross-links (DPCs). In the present work, a mass spectrometry-based proteomics approach was employed to characterize PM- and NOR-mediated DNA–Protein Cross-Linking in human cells. Following treatment of human fibrosarcoma cells (HT1080) with cytotoxic concentrations of PM, over 130 Proteins were found to be covalently trapped to DNA, including those involved in transcriptional regulation, RNA splicing/processing, chromatin organization, and Protein transport. HPLC-ESI+-MS/MS analysis of proteolytic digests of DPC-containing DNA from NOR-treated cells revealed a con...
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mechlorethamine induced dna Protein Cross Linking in human fibrosarcoma ht1080 cells
Journal of Proteome Research, 2011Co-Authors: Erin D Michaelsonrichie, Simona G Codreanu, Rachel Loeber, Colin R Campbell, Daniel C. Liebler, Xun Ming, Natalia Y TretyakovaAbstract:Antitumor nitrogen mustards, such as bis(2-chloroethyl)methylamine (mechlorethamine), are useful chemotherapeutic agents with a long history of clinical application. The antitumor effects of nitrogen mustards are attributed to their ability to induce DNA–DNA and DNA–Protein Cross-links (DPCs) that block DNA replication. In the present work, a mass spectrometry-based methodology was employed to characterize in vivo DNA–Protein Cross-Linking following treatment of human fibrosarcoma (HT1080) cells with cytotoxic concentrations of mechlorethamine. A combination of mass spectrometry-based proteomics and immunological detection was used to identify 38 nuclear Proteins that were covalently Cross-linked to chromosomal DNA following treatment with mechlorethamine. Isotope dilution HPLC–ESI+–MS/MS analysis of total proteolytic digests revealed a concentration-dependent formation of N-[2-(S-cysteinyl)ethyl]-N-[2-(guan-7-yl)ethyl]methylamine (Cys-N7G-EMA) conjugates, indicating that mechlorethamine Cross-links cyste...
Colin R Campbell - One of the best experts on this subject based on the ideXlab platform.
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mass spectrometry based proteomics study of cisplatin induced dna Protein Cross Linking in human fibrosarcoma ht1080 cells
Chemical Research in Toxicology, 2017Co-Authors: Xun Ming, Arnold Groehler, Erin D Michaelsonrichie, Peter W Villalta, Colin R Campbell, Natalia Y TretyakovaAbstract:Platinum-based antitumor drugs such as 1,1,2,2-cis-diamminedichloroplatinum(II) (cisplatin), carboplatin, and oxaliplatin are currently used to treat nearly 50% of all cancer cases, and novel platinum based agents are under development. The antitumor effects of cisplatin and other platinum compounds are attributed to their ability to induce interstrand DNA–DNA Cross-links, which are thought to inhibit tumor cell growth by blocking DNA replication and/or preventing transcription. However, platinum agents also induce significant numbers of unusually bulky and helix-distorting DNA–Protein Cross-links (DPCs), which are poorly characterized because of their unusual complexity. We and others have previously shown that DPCs block DNA replication and transcription and causes toxicity in human cells, potentially contributing to the biological effects of platinum agents. In the present work, we have undertaken a system-wide investigation of cisplatin-mediated DNA–Protein Cross-Linking in human fibrosarcoma (HT1080)...
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covalent dna Protein Cross Linking by phosphoramide mustard and nornitrogen mustard in human cells
Chemical Research in Toxicology, 2016Co-Authors: Arnold Groehler, Peter W Villalta, Colin R Campbell, Natalia Y TretyakovaAbstract:N,N-Bis-(2-chloroethyl)-phosphorodiamidic acid (phosphoramide mustard, PM) and N,N-bis-(2-chloroethyl)-amine (nornitrogen mustard, NOR) are the two biologically active metabolites of cyclophosphamide, a DNA alkylating drug commonly used to treat lymphomas, breast cancer, certain brain cancers, and autoimmune diseases. PM and NOR are reactive bis-electrophiles capable of Cross-Linking cellular biomolecules to form covalent DNA–DNA and DNA–Protein Cross-links (DPCs). In the present work, a mass spectrometry-based proteomics approach was employed to characterize PM- and NOR-mediated DNA–Protein Cross-Linking in human cells. Following treatment of human fibrosarcoma cells (HT1080) with cytotoxic concentrations of PM, over 130 Proteins were found to be covalently trapped to DNA, including those involved in transcriptional regulation, RNA splicing/processing, chromatin organization, and Protein transport. HPLC-ESI+-MS/MS analysis of proteolytic digests of DPC-containing DNA from NOR-treated cells revealed a con...
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mechlorethamine induced dna Protein Cross Linking in human fibrosarcoma ht1080 cells
Journal of Proteome Research, 2011Co-Authors: Erin D Michaelsonrichie, Simona G Codreanu, Rachel Loeber, Colin R Campbell, Daniel C. Liebler, Xun Ming, Natalia Y TretyakovaAbstract:Antitumor nitrogen mustards, such as bis(2-chloroethyl)methylamine (mechlorethamine), are useful chemotherapeutic agents with a long history of clinical application. The antitumor effects of nitrogen mustards are attributed to their ability to induce DNA–DNA and DNA–Protein Cross-links (DPCs) that block DNA replication. In the present work, a mass spectrometry-based methodology was employed to characterize in vivo DNA–Protein Cross-Linking following treatment of human fibrosarcoma (HT1080) cells with cytotoxic concentrations of mechlorethamine. A combination of mass spectrometry-based proteomics and immunological detection was used to identify 38 nuclear Proteins that were covalently Cross-linked to chromosomal DNA following treatment with mechlorethamine. Isotope dilution HPLC–ESI+–MS/MS analysis of total proteolytic digests revealed a concentration-dependent formation of N-[2-(S-cysteinyl)ethyl]-N-[2-(guan-7-yl)ethyl]methylamine (Cys-N7G-EMA) conjugates, indicating that mechlorethamine Cross-links cyste...
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DNA-Protein Cross-Linking by 1,2,3,4-diepoxybutane.
Journal of proteome research, 2010Co-Authors: Erin D. Michaelson-richie, Simona G Codreanu, Rachel Loeber, Colin R Campbell, Daniel C. Liebler, Xun Ming, Natalia Y TretyakovaAbstract:1,2,3,4-Diepoxybutane (DEB) is a strongly genotoxic diepoxide hypothesized to be the ultimate carcinogenic metabolite of the common industrial chemical and environmental carcinogen 1,3-butadiene. D...
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Proteomic Analysis of DNA-Protein Cross-Linking by Antitumor Nitrogen Mustards
Chemical research in toxicology, 2009Co-Authors: Rachel Loeber, Simona G Codreanu, Erin D. Michaelson-richie, Colin R Campbell, Daniel C. Liebler, Natalia Y TretyakovaAbstract:Nitrogen mustards are antitumor agents used clinically for the treatment of a variety of neoplastic conditions. The biological activity of these compounds is typically attributed to their ability to induce DNA−DNA Cross-links. However, nitrogen mustards are able to produce a variety of other lesions, including DNA−Protein Cross-links (DPCs). DPCs induced by nitrogen mustards are not well-characterized because of their structural complexity and the insufficient specificity and sensitivity of previously available experimental methodologies. In the present work, affinity capture methodology in combination with mass spectrometry-based proteomics was employed to identify mammalian Proteins that form covalent Cross-links to DNA in the presence of a simple nitrogen mustard, mechlorethamine. Following incubation of 5′-biotinylated DNA duplexes with nuclear Protein extracts, DPCs were isolated by affinity capture on streptavidin beads, and the Cross-linked Proteins were identified by high-performance liquid chroma...
Xun Ming - One of the best experts on this subject based on the ideXlab platform.
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mass spectrometry based proteomics study of cisplatin induced dna Protein Cross Linking in human fibrosarcoma ht1080 cells
Chemical Research in Toxicology, 2017Co-Authors: Xun Ming, Arnold Groehler, Erin D Michaelsonrichie, Peter W Villalta, Colin R Campbell, Natalia Y TretyakovaAbstract:Platinum-based antitumor drugs such as 1,1,2,2-cis-diamminedichloroplatinum(II) (cisplatin), carboplatin, and oxaliplatin are currently used to treat nearly 50% of all cancer cases, and novel platinum based agents are under development. The antitumor effects of cisplatin and other platinum compounds are attributed to their ability to induce interstrand DNA–DNA Cross-links, which are thought to inhibit tumor cell growth by blocking DNA replication and/or preventing transcription. However, platinum agents also induce significant numbers of unusually bulky and helix-distorting DNA–Protein Cross-links (DPCs), which are poorly characterized because of their unusual complexity. We and others have previously shown that DPCs block DNA replication and transcription and causes toxicity in human cells, potentially contributing to the biological effects of platinum agents. In the present work, we have undertaken a system-wide investigation of cisplatin-mediated DNA–Protein Cross-Linking in human fibrosarcoma (HT1080)...
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1,2,3,4-Diepoxybutane-induced DNA-Protein Cross-Linking in human fibrosarcoma (HT1080) cells
Journal of proteome research, 2013Co-Authors: Teshome B. Gherezghiher, Xun Ming, Peter W VillaltaAbstract:1,2,3,4-Diepoxybutane (DEB) is the key carcinogenic metabolite of 1,3-butadiene (BD), an important industrial and environmental chemical present in urban air and in cigarette smoke. DEB is a genotoxic bis-electrophile capable of Cross-Linking cellular biomolecules to form DNA–DNA and DNA–Protein Cross-links (DPCs). In the present work, mass spectrometry-based proteomics was employed to characterize DEB-mediated DNA–Protein Cross-Linking in human fibrosarcoma (HT1080) cells. Over 150 Proteins including histones, high mobility group Proteins, transcription factors, splicing factors, and tubulins were found among those covalently Cross-linked to chromosomal DNA in the presence of DEB. A large portion of the Cross-linked Proteins are known factors involved in DNA binding, transcriptional regulation, cell signaling, DNA repair, and DNA damage response. HPLC–ESI+–MS/MS analysis of total proteolytic digests revealed the presence of 1-(S-cysteinyl)-4-(guan-7-yl)-2,3-butanediol conjugates, confirming that DEB form...
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mechlorethamine induced dna Protein Cross Linking in human fibrosarcoma ht1080 cells
Journal of Proteome Research, 2011Co-Authors: Erin D Michaelsonrichie, Simona G Codreanu, Rachel Loeber, Colin R Campbell, Daniel C. Liebler, Xun Ming, Natalia Y TretyakovaAbstract:Antitumor nitrogen mustards, such as bis(2-chloroethyl)methylamine (mechlorethamine), are useful chemotherapeutic agents with a long history of clinical application. The antitumor effects of nitrogen mustards are attributed to their ability to induce DNA–DNA and DNA–Protein Cross-links (DPCs) that block DNA replication. In the present work, a mass spectrometry-based methodology was employed to characterize in vivo DNA–Protein Cross-Linking following treatment of human fibrosarcoma (HT1080) cells with cytotoxic concentrations of mechlorethamine. A combination of mass spectrometry-based proteomics and immunological detection was used to identify 38 nuclear Proteins that were covalently Cross-linked to chromosomal DNA following treatment with mechlorethamine. Isotope dilution HPLC–ESI+–MS/MS analysis of total proteolytic digests revealed a concentration-dependent formation of N-[2-(S-cysteinyl)ethyl]-N-[2-(guan-7-yl)ethyl]methylamine (Cys-N7G-EMA) conjugates, indicating that mechlorethamine Cross-links cyste...
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DNA-Protein Cross-Linking by 1,2,3,4-diepoxybutane.
Journal of proteome research, 2010Co-Authors: Erin D. Michaelson-richie, Simona G Codreanu, Rachel Loeber, Colin R Campbell, Daniel C. Liebler, Xun Ming, Natalia Y TretyakovaAbstract:1,2,3,4-Diepoxybutane (DEB) is a strongly genotoxic diepoxide hypothesized to be the ultimate carcinogenic metabolite of the common industrial chemical and environmental carcinogen 1,3-butadiene. D...
Peter W Villalta - One of the best experts on this subject based on the ideXlab platform.
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mass spectrometry based proteomics study of cisplatin induced dna Protein Cross Linking in human fibrosarcoma ht1080 cells
Chemical Research in Toxicology, 2017Co-Authors: Xun Ming, Arnold Groehler, Erin D Michaelsonrichie, Peter W Villalta, Colin R Campbell, Natalia Y TretyakovaAbstract:Platinum-based antitumor drugs such as 1,1,2,2-cis-diamminedichloroplatinum(II) (cisplatin), carboplatin, and oxaliplatin are currently used to treat nearly 50% of all cancer cases, and novel platinum based agents are under development. The antitumor effects of cisplatin and other platinum compounds are attributed to their ability to induce interstrand DNA–DNA Cross-links, which are thought to inhibit tumor cell growth by blocking DNA replication and/or preventing transcription. However, platinum agents also induce significant numbers of unusually bulky and helix-distorting DNA–Protein Cross-links (DPCs), which are poorly characterized because of their unusual complexity. We and others have previously shown that DPCs block DNA replication and transcription and causes toxicity in human cells, potentially contributing to the biological effects of platinum agents. In the present work, we have undertaken a system-wide investigation of cisplatin-mediated DNA–Protein Cross-Linking in human fibrosarcoma (HT1080)...
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covalent dna Protein Cross Linking by phosphoramide mustard and nornitrogen mustard in human cells
Chemical Research in Toxicology, 2016Co-Authors: Arnold Groehler, Peter W Villalta, Colin R Campbell, Natalia Y TretyakovaAbstract:N,N-Bis-(2-chloroethyl)-phosphorodiamidic acid (phosphoramide mustard, PM) and N,N-bis-(2-chloroethyl)-amine (nornitrogen mustard, NOR) are the two biologically active metabolites of cyclophosphamide, a DNA alkylating drug commonly used to treat lymphomas, breast cancer, certain brain cancers, and autoimmune diseases. PM and NOR are reactive bis-electrophiles capable of Cross-Linking cellular biomolecules to form covalent DNA–DNA and DNA–Protein Cross-links (DPCs). In the present work, a mass spectrometry-based proteomics approach was employed to characterize PM- and NOR-mediated DNA–Protein Cross-Linking in human cells. Following treatment of human fibrosarcoma cells (HT1080) with cytotoxic concentrations of PM, over 130 Proteins were found to be covalently trapped to DNA, including those involved in transcriptional regulation, RNA splicing/processing, chromatin organization, and Protein transport. HPLC-ESI+-MS/MS analysis of proteolytic digests of DPC-containing DNA from NOR-treated cells revealed a con...
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Covalent DNA–Protein Cross-Linking by Phosphoramide Mustard and Nornitrogen Mustard in Human Cells
2016Co-Authors: Arnold Groehler, Peter W Villalta, Colin Campbell, Natalia TretyakovaAbstract:N,N-Bis-(2-chloroethyl)-phosphorodiamidic acid (phosphoramide mustard, PM) and N,N-bis-(2-chloroethyl)-amine (nornitrogen mustard, NOR) are the two biologically active metabolites of cyclophosphamide, a DNA alkylating drug commonly used to treat lymphomas, breast cancer, certain brain cancers, and autoimmune diseases. PM and NOR are reactive bis-electrophiles capable of Cross-Linking cellular biomolecules to form covalent DNA–DNA and DNA–Protein Cross-links (DPCs). In the present work, a mass spectrometry-based proteomics approach was employed to characterize PM- and NOR-mediated DNA–Protein Cross-Linking in human cells. Following treatment of human fibrosarcoma cells (HT1080) with cytotoxic concentrations of PM, over 130 Proteins were found to be covalently trapped to DNA, including those involved in transcriptional regulation, RNA splicing/processing, chromatin organization, and Protein transport. HPLC-ESI+-MS/MS analysis of proteolytic digests of DPC-containing DNA from NOR-treated cells revealed a concentration-dependent formation of N-[2-[cysteinyl]ethyl]-N-[2-(guan-7-yl)ethyl]amine (Cys-NOR-N7G) conjugates, confirming that it Cross-links cysteine thiols of Proteins to the N7 position of guanines in DNA. Cys-NOR-N7G adduct numbers were higher in NER-deficient xeroderma pigmentosum cells (XPA) as compared with repair proficient cells. Furthermore, both XPA and FANCD2 deficient cells were sensitized to PM treatment as compared to that of wild type cells, suggesting that Fanconi anemia and nucleotide excision repair pathways are involved in the removal of cyclophosphamide-induced DNA damage
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1,2,3,4-Diepoxybutane-induced DNA-Protein Cross-Linking in human fibrosarcoma (HT1080) cells
Journal of proteome research, 2013Co-Authors: Teshome B. Gherezghiher, Xun Ming, Peter W VillaltaAbstract:1,2,3,4-Diepoxybutane (DEB) is the key carcinogenic metabolite of 1,3-butadiene (BD), an important industrial and environmental chemical present in urban air and in cigarette smoke. DEB is a genotoxic bis-electrophile capable of Cross-Linking cellular biomolecules to form DNA–DNA and DNA–Protein Cross-links (DPCs). In the present work, mass spectrometry-based proteomics was employed to characterize DEB-mediated DNA–Protein Cross-Linking in human fibrosarcoma (HT1080) cells. Over 150 Proteins including histones, high mobility group Proteins, transcription factors, splicing factors, and tubulins were found among those covalently Cross-linked to chromosomal DNA in the presence of DEB. A large portion of the Cross-linked Proteins are known factors involved in DNA binding, transcriptional regulation, cell signaling, DNA repair, and DNA damage response. HPLC–ESI+–MS/MS analysis of total proteolytic digests revealed the presence of 1-(S-cysteinyl)-4-(guan-7-yl)-2,3-butanediol conjugates, confirming that DEB form...
Rachel Loeber - One of the best experts on this subject based on the ideXlab platform.
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mechlorethamine induced dna Protein Cross Linking in human fibrosarcoma ht1080 cells
Journal of Proteome Research, 2011Co-Authors: Erin D Michaelsonrichie, Simona G Codreanu, Rachel Loeber, Colin R Campbell, Daniel C. Liebler, Xun Ming, Natalia Y TretyakovaAbstract:Antitumor nitrogen mustards, such as bis(2-chloroethyl)methylamine (mechlorethamine), are useful chemotherapeutic agents with a long history of clinical application. The antitumor effects of nitrogen mustards are attributed to their ability to induce DNA–DNA and DNA–Protein Cross-links (DPCs) that block DNA replication. In the present work, a mass spectrometry-based methodology was employed to characterize in vivo DNA–Protein Cross-Linking following treatment of human fibrosarcoma (HT1080) cells with cytotoxic concentrations of mechlorethamine. A combination of mass spectrometry-based proteomics and immunological detection was used to identify 38 nuclear Proteins that were covalently Cross-linked to chromosomal DNA following treatment with mechlorethamine. Isotope dilution HPLC–ESI+–MS/MS analysis of total proteolytic digests revealed a concentration-dependent formation of N-[2-(S-cysteinyl)ethyl]-N-[2-(guan-7-yl)ethyl]methylamine (Cys-N7G-EMA) conjugates, indicating that mechlorethamine Cross-links cyste...
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DNA-Protein Cross-Linking by 1,2,3,4-diepoxybutane.
Journal of proteome research, 2010Co-Authors: Erin D. Michaelson-richie, Simona G Codreanu, Rachel Loeber, Colin R Campbell, Daniel C. Liebler, Xun Ming, Natalia Y TretyakovaAbstract:1,2,3,4-Diepoxybutane (DEB) is a strongly genotoxic diepoxide hypothesized to be the ultimate carcinogenic metabolite of the common industrial chemical and environmental carcinogen 1,3-butadiene. D...
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Proteomic Analysis of DNA-Protein Cross-Linking by Antitumor Nitrogen Mustards
Chemical research in toxicology, 2009Co-Authors: Rachel Loeber, Simona G Codreanu, Erin D. Michaelson-richie, Colin R Campbell, Daniel C. Liebler, Natalia Y TretyakovaAbstract:Nitrogen mustards are antitumor agents used clinically for the treatment of a variety of neoplastic conditions. The biological activity of these compounds is typically attributed to their ability to induce DNA−DNA Cross-links. However, nitrogen mustards are able to produce a variety of other lesions, including DNA−Protein Cross-links (DPCs). DPCs induced by nitrogen mustards are not well-characterized because of their structural complexity and the insufficient specificity and sensitivity of previously available experimental methodologies. In the present work, affinity capture methodology in combination with mass spectrometry-based proteomics was employed to identify mammalian Proteins that form covalent Cross-links to DNA in the presence of a simple nitrogen mustard, mechlorethamine. Following incubation of 5′-biotinylated DNA duplexes with nuclear Protein extracts, DPCs were isolated by affinity capture on streptavidin beads, and the Cross-linked Proteins were identified by high-performance liquid chroma...
