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• W2324347180 endingPage "233" @default.
• W2324347180 startingPage "224" @default.
• W2324347180 abstract "O(6)-alkylguanine-DNA alkyltransferases (AGT) are responsible for the removal of alkylation at both the O(6) atom of guanine and O(4) atom of thymine. AGT homologues show vast substrate differences with respect to the size of the adduct and which alkylated atoms they can restore. The human AGT (hAGT) has poor capabilities for removal of methylation at the O(4) atom of thymidine, which is not the case in most homologues. No structural data are available to explain this poor hAGT repair. We prepared and characterized O(6)G-butylene-O(4)T (XLGT4) and O(6)G-heptylene-O(4)T (XLGT7) interstrand cross-linked (ICL) DNA as probes for hAGT and the Escherichia coli homologues, OGT and Ada-C, for the formation of DNA-AGT covalent complexes. XLGT7 reacted only with hAGT and did so with a cross-linking efficiency of 25%, while XLGT4 was inert to all AGT tested. The hAGT mediated repair of XLGT7 occurred slowly, on the order of hours as opposed to the repair of O(6)-methyl-2'-deoxyguanosine which requires seconds. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of the repair reaction revealed the formation of a covalent complex with an observed migration in accordance with a DNA-AGT complex. The identity of this covalent complex, as determined by mass spectrometry, was composed of a heptamethylene bridge between the O(4) atom of thymidine (in an 11-mer DNA strand) to residue Cys145 of hAGT. This procedure can be applied to produce well-defined covalent complexes between AGT with DNA." @default.
• W2324347180 created "2016-06-24" @default.
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• W2324347180 creator A5056842088 @default.
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• W2324347180 date "2013-01-24" @default.
• W2324347180 modified "2023-09-23" @default.
• W2324347180 title "Preparation of Covalently Linked Complexes Between DNA and O6-Alkylguanine-DNA Alkyltransferase Using Interstrand Cross-Linked DNA" @default.
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• W2324347180 doi "https://doi.org/10.1021/bc300553u" @default.
• W2324347180 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/23347328" @default.
• W2324347180 hasPublicationYear "2013" @default.
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