title: Molecular recognition of T:G mismatched base pairs in DNA as studied by electrospray ionization mass spectrometry.
abstract: Postreplicative mismatch repair (MMR) is a cellular system involved in the recognition and correction of DNA polymerase errors that escape detection in proofreading. Of the various mismatched bases, T:G pairing in DNA is one of the more common mutations leading to the formation of tumors in humans. In addition, the absence of the MMR system can generate resistance to several chemotherapeutic agents, particularly DNA-damaging substances. The main purpose of this study was the setup and validation of an electrospray ionization (ESI) mass spectrometry method for the identification of small molecules that are able to recognize T:G mismatches in DNA targets. These findings could be useful for the discovery of new antitumor drugs. The analytical method is based on the ability of electrospray to preserve the noncovalent adducts present in solution and transfer them to the gas phase. Lexitropsin derivatives (polyimidazole compounds) have been previously described as selective for T:G mismatch binding by NMR and ITC studies. We synthesized and tested various polyimidazole derivatives, one of which in particular (NMS-057) showed a higher affinity for an oligonucleotide DNA sequence containing a T:G mismatched base pair. To rationalize these findings, molecular docking studies were performed using available NMR structures. Moreover, ESI-MS experiments, performed on an orbitrap mass spectrometer, highlighted the formation of heterodimeric complexes between DNA sequences, distamycin A, and polyimidazole compounds. Our results confirm that this ESI method could be a valuable tool for the identification of new molecules able to specifically recognize T:G mismatched base pairs.
authors.affiliation: Oncology Business Unit, Nerviano Medical Sciences, Viale Pasteur 10, 20014 Nerviano (MI), Italy. federico.riccardi@nervianoms.com
authors.forename: Federico
authors.initials: F
authors.lastname: Riccardi Sirtori
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Aldini
authors.forename: Maristella
authors.initials: M
authors.lastname: Colombo
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Colombo
authors.forename: Jan
authors.initials: J
authors.lastname: Malyszko
authors.forename: Giulio
authors.initials: G
authors.lastname: Vistoli
authors.forename: Roberto
authors.initials: R
authors.lastname: D'Alessio
mesh_terms: D020137:Base Pair Mismatch; D004247:DNA / Q000737:chemistry*; D006147:Guanine / Q000737:chemistry*; D009429:Netropsin / Q000031:analogs & derivatives / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013941:Thymine / Q000737:chemistry*
chemical_list: C053654:lexitropsin; D006147:Guanine; D009429:Netropsin; D004247:DNA; D013941:Thymine