Characterization of nucleic acid higher order structure by high-resolution tandem mass spectrometry |
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Authors: | Jingjie Mo Kristina Håkansson |
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Institution: | (1) Department of Chemistry, University of Michigan, 930 North University Avenue, Ann Arbor, MI 48109-1055, USA |
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Abstract: | Mass spectrometry (MS) is extensively used for the identification and sequencing of nucleic acids but has so far seen limited
use for characterization of their higher order structures. Here, we have applied a range of different tandem mass spectrometry
techniques, including electron detachment dissociation (EDD), infrared multiphoton dissociation (IRMPD), activated ion (AI)
EDD, and EDD/IRMPD MS3, in a Fourier transform ion cyclotron resonance mass spectrometer to the characterization of three isomeric 15mer DNAs with
different sequences and predicted solution-phase structures. Our goal was to explore whether their structural differences
could be directly probed with these techniques. We found that all three 15mers had higher order structures in the gas phase,
although preferred structures were predicted for only two of them in solution. Nevertheless, EDD, AI EDD, and EDD/IRMPD MS3 experiments yielded different cleavage patterns with less backbone fragmentation for the more stable solution-phase structure
than for the other two 15mers. By contrast, no major differences were observed in IRMPD, although the extent of backbone cleavage
was higher with that technique for all three 15mers. Thus, experiments utilizing the radical ion chemistry of EDD can provide
complementary structural information compared to traditional slow heating methods, such as IRMPD, for structured nucleic acids. |
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Keywords: | Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) Infrared mulitphoton dissociation (IRMPD) Electron detachment dissociation (EDD) Oligonucleotide Nucleic acid DNA |
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