[PMID]: | 26692166 |
[Au] Autor: | Groehler A; Villalta PW; Campbell C; Tretyakova N |
[Ad] Endereço: | Department of Medicinal Chemistry, ‡Department of Pharmacology, and §Masonic Cancer Center, University of Minnesota , Minneapolis, Minnesota 55455, United States. |
[Ti] Título: | Covalent DNA-Protein Cross-Linking by Phosphoramide Mustard and Nornitrogen Mustard in Human Cells. |
[So] Source: | Chem Res Toxicol;29(2):190-202, 2016 Feb 15. |
[Is] ISSN: | 1520-5010 |
[Cp] País de publicação: | United States |
[La] Idioma: | eng |
[Ab] Resumo: | 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. |
[Mh] Termos MeSH primário: |
Alquilantes/química DNA/química Compostos de Mostarda Nitrogenada/química Mostardas de Fosforamida/química Proteínas/química
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[Mh] Termos MeSH secundário: |
Linhagem Celular Tumoral Cromatografia Líquida de Alta Pressão DNA/metabolismo Adutos de DNA/análise Seres Humanos Peptídeos/análise Proteínas/metabolismo Proteômica Espectrometria de Massas por Ionização por Electrospray
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[Pt] Tipo de publicação: | JOURNAL ARTICLE; RESEARCH SUPPORT, N.I.H., EXTRAMURAL |
[Nm] Nome de substância:
| 0 (Alkylating Agents); 0 (DNA Adducts); 0 (Nitrogen Mustard Compounds); 0 (Peptides); 0 (Phosphoramide Mustards); 0 (Proteins); 10159-53-2 (phosphoramide mustard); 9007-49-2 (DNA); IHV3I38UXH (nornitrogen mustard) |
[Em] Mês de entrada: | 1610 |
[Cu] Atualização por classe: | 170220 |
[Lr] Data última revisão:
| 170220 |
[Sb] Subgrupo de revista: | IM |
[Da] Data de entrada para processamento: | 151223 |
[St] Status: | MEDLINE |
[do] DOI: | 10.1021/acs.chemrestox.5b00430 |
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