Analysis of platinum adducts with DNA nucleotides and nucleosides by capillary electrophoresis coupled to ESI-MS: indications of guanosine 5'-monophosphate O6-N7 chelation

Chembiochem. 2004 Nov 5;5(11):1543-9. doi: 10.1002/cbic.200400015.

Abstract

DNA is the ultimate target of platinum-based anticancer therapy. Since the N7 of guanine is known to be the major binding site of cisplatin and its analogues, adduct formation with model nucleotides, especially 2'-deoxyguanosine 5'-monophosphate (dGMP), has been studied in detail. During the last few years a coupled capillary eletrophoresis/electrospray-ionization mass spectrometry (CE/ESI-MS) method has been advantageously used in order to separate and identify platinum adducts with nucleotides in submillimolar concentrations in aqueous solutions. Beside the bisadduct, [Pt(NH(3))(2)(dNMP)(2)](2-) (NMP=2'-deoxynucleoside 5'-monophosphate), and the well-known monochloro and monohydroxo adducts, [Pt(NH(3))(2)Cl(dNMP)](-) and [Pt(NH(3))(2)(dNMP)OH](-), respectively, a third kind of monoadduct species with a composition of [Pt(NH(3))(2)(dNMP)](-) can be separated by CE and detected through the m/z values measured with ESI-MS. Different experimental setups indicate the existence of an O(6)-N7 chelate, whereas the formation of N7-alphaPO(4) macrochelates or dinuclear species is unlikely. Additionally, offline MS experiments with 2'-deoxyguanosine (dG) and stabilization of the controversially discussed O(6)-N7 chelate by oxidation with hydrogen peroxide support the assumption of the existence of O(6)-N7 chelation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cisplatin / chemistry*
  • DNA Adducts / analysis
  • DNA Adducts / chemistry*
  • Deoxyguanine Nucleotides / chemistry
  • Electrophoresis, Capillary / methods
  • Guanosine Monophosphate / chemistry*
  • Inosine Monophosphate / analogs & derivatives*
  • Inosine Monophosphate / chemistry
  • Molecular Structure
  • Nucleosides / chemistry*
  • Nucleotides / chemistry*
  • Organoplatinum Compounds / chemistry*
  • Spectrometry, Mass, Electrospray Ionization / methods

Substances

  • DNA Adducts
  • Deoxyguanine Nucleotides
  • Nucleosides
  • Nucleotides
  • Organoplatinum Compounds
  • Inosine Monophosphate
  • deoxyinosine monophosphate
  • 2'-deoxyguanosine 5'-phosphate
  • Guanosine Monophosphate
  • Cisplatin