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RESEARCH FRONT
Contents Vol 64(6)

Developments in Electrospray Ionization Mass Spectrometry of Non-Covalent DNA–Ligand Complexes

Jennifer L. Beck A
+ Author Affiliations
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A School of Chemistry, University of Wollongong, NSW 2522, Australia. Email: jbeck@uow.edu.au




Jennifer's undergraduate and Ph.D. studies were carried out at the University of Queensland. Her Ph.D. was titled ‘Metal Ion Binding Sites of Purple Phosphatases’. The theme of biological chemistry was continued through her postdoctoral appointments: first at the University of Illinois at Urbana-Champaign (electron transfer pathways between cytochrome P-450cam and putidaredoxin) and later at the ANU (protein subunits of DNA polymerase III of E. coli). After a substantial career interruption, she re-entered science in the new field of biomolecular mass spectrometry at the University of Wollongong. Her current work involves applications of ESI-MS and other biophysical techniques to investigate non-covalent complexes of the E. coli replisome, DNA–ligand complexes, and new work involves qDNA-associated proteins.

Australian Journal of Chemistry 64(6) 705-717 https://doi.org/10.1071/CH11046
Submitted: 26 January 2011  Accepted: 13 April 2011   Published: 27 June 2011

Abstract

Many anti-cancer drugs function by binding non-covalently to double-stranded (ds) DNA. Electrospray ionization mass spectrometry (ESI-MS) has emerged over the past decade as a sensitive technique for the determination of stoichiometries and relative binding affinities of DNA–ligand interactions. The chromosome contains nucleotide sequences, for example, guanosine-rich regions, that predispose them to the formation of higher order structures such as quadruplex DNA (qDNA). Sequences that form qDNA are found in the telomeres. The proposal that ligands that stabilize qDNA might interfere with the activity of telomerase in cancer cells has stimulated the search for ligands that are selective for qDNA over dsDNA. The insights gained from the development of ESI-MS methods for analysis of non-covalent dsDNA–ligand complexes are now being applied in the search for qDNA-selective ligands. ESI-MS is a useful first-pass screening technique for qDNA-binding ligands. This short review describes some experimental considerations for ESI-MS analysis of DNA–ligand complexes, briefly addresses the question of whether non-covalent DNA–ligand complexes are faithfully transferred from solution to the gas phase, discusses ion mobility mass spectrometry as a technique for probing this issue, and highlights some recent ESI-MS studies of qDNA-selective ligands.


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