BINDING-SPECIFICITY OF POST-ACTIVATED NEOCARZINOSTATIN CHROMOPHORE DRUG-BULGED DNA COMPLEX STUDIED USING ELECTROSPRAY-IONIZATION MASS-SPECTROMETRY

Electrospray ionization mass spectrometry (ESI-MS) was employed to cha racterize the binding specificity of a bulged 22-mer DNA hairpin with a post-activated neocarzinostatin chromophore (NCS-Chrom) having two s imilar forms, where 2a has an H in a location for which 2b has it repl aced by a CH2OH group, Specific binding of 2a to the bulged 22-mer DNA was observed whereas little binding was detected for 2a to non-bulged DNA 19-mer and 12-mer duplexes. The stoichiometry of the 22-mer DNA c omplex with 2a was determined to be predominantly 1:1. Substitution of hydrogen in 2a for the hydroxymethylene group in 2b dramatically redu ced the binding strength to the 22-mer DNA. Little complex formation w as observed for 2b and 22-mer DNA based upon the ESI-MS data, consiste nt with earlier fluorescence studies, The results indicate that ESI-MS can be a sensitive technique for probing conformational specificity i n studies of biomolecular binding.