title: The in vitro methylation of DNA by a minor groove binding methyl sulfonate ester.
abstract: The preparation of sequence and groove specific DNA methylating agents based on N-methylpyrrolecarboxamide subunits appended with an O-methyl sulfonate ester functionality (MeOSO2(CH2)2-Lex) has previously been described [Zhang, Y., Chen, F.-X., Mehta, P., and Gold, B. (1993) Biochemistry 32,7954-7965]. In contrast to simple methyl sulfonate esters, e.g., methyl methanesulfonate (MMS), which predominantly methylate at 7-guanine, MeOSO2-(CH2)2-Lex affords N3-methyladenine (3-MeAde) as its major adduct. Using competitive ELISA determinations, the methylation at major and minor groove sites in calf thymus DNA by MeOSO2(CH2)2-Lex has been precisely quantitated. The yields of N7-methylguanine (7-MeGua), 3-MeAde, and O6-methyldeoxyguanosine (6-Me-dGuo) are 0.424, 3.195, and 0.0027 mmol of adduct/mol of DNA, respectively, using 10 microM MeOSO2(CH2)2-Lex and 100 microM DNA. This compares to 0.773, 0.072, and 0.0033 mmol of adduct/mol of DNA for 7-MeGua, 3-MeAde, and 6-Me-dGuo, respectively, using MMS. The increase in the yield of 3-MeAde due to the minor groove equilibrium binding properties of MeOSO2(CH2)2-Lex is approximately 40-fold relative to MMS.
authors.affiliation: Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska Medical Center, Omaha 68198-6805, USA.
authors.forename: L
authors.initials: L
authors.lastname: Encell
authors.forename: D E
authors.initials: DE
authors.lastname: Shuker
authors.forename: P G
authors.initials: PG
authors.lastname: Foiles
authors.forename: B
authors.initials: B
authors.lastname: Gold
mesh_terms: D000225:Adenine / Q000031:analogs & derivatives* / Q000032:analysis / Q000096:biosynthesis; D000476:Alkanesulfonates / Q000378:metabolism*; D000818:Animals; D000970:Antineoplastic Agents; D001665:Binding Sites; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism*; D018736:DNA Adducts / Q000032:analysis* / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006147:Guanine / Q000031:analogs & derivatives* / Q000032:analysis / Q000096:biosynthesis; D006151:Guanosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000096:biosynthesis; D066298:In Vitro Techniques; D008741:Methyl Methanesulfonate / Q000378:metabolism; D008745:Methylation; D009429:Netropsin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D011758:Pyrroles / Q000378:metabolism*
chemical_list: D000476:Alkanesulfonates; D000970:Antineoplastic Agents; D018736:DNA Adducts; D011758:Pyrroles; C053654:lexitropsin; D006151:Guanosine; C101157:1-methyl-4-(((1-methyl-4-((3-((methylsulfonyl)oxy)-1-oxopropyl)amino)-1H-pyrrol-2-yl)carbonyl)amino)-N-propyl-1H-pyrrole-2-carboxamide; C025946:3-methyladenine; D006147:Guanine; D009429:Netropsin; C008450:7-methylguanine; C046175:6'-O-methylguanosine; D004247:DNA; D008741:Methyl Methanesulfonate; D000225:Adenine