The in vitro methylation of DNA by a minor groove binding methyl sulfonate ester

Chem Res Toxicol. Apr-May 1996;9(3):563-7. doi: 10.1021/tx9501849.

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenine / analogs & derivatives*
  • Adenine / analysis
  • Adenine / biosynthesis
  • Alkanesulfonates / metabolism*
  • Animals
  • Antineoplastic Agents
  • Binding Sites
  • Cattle
  • Chromatography, High Pressure Liquid
  • DNA / metabolism*
  • DNA Adducts / analysis*
  • DNA Adducts / biosynthesis
  • Dose-Response Relationship, Drug
  • Enzyme-Linked Immunosorbent Assay
  • Guanine / analogs & derivatives*
  • Guanine / analysis
  • Guanine / biosynthesis
  • Guanosine / analogs & derivatives*
  • Guanosine / analysis
  • Guanosine / biosynthesis
  • In Vitro Techniques
  • Methyl Methanesulfonate / metabolism
  • Methylation
  • Netropsin / analogs & derivatives*
  • Netropsin / chemistry
  • Netropsin / metabolism
  • Pyrroles / metabolism*

Substances

  • Alkanesulfonates
  • Antineoplastic Agents
  • DNA Adducts
  • Pyrroles
  • lexitropsin
  • Guanosine
  • 1-methyl-4-(((1-methyl-4-((3-((methylsulfonyl)oxy)-1-oxopropyl)amino)-1H-pyrrol-2-yl)carbonyl)amino)-N-propyl-1H-pyrrole-2-carboxamide
  • 3-methyladenine
  • Guanine
  • Netropsin
  • 7-methylguanine
  • 6'-O-methylguanosine
  • DNA
  • Methyl Methanesulfonate
  • Adenine