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PubMed · 10746015

Affinity NMR: decoding DNA binding.

R C Anderson·M Lin·M J Shapiro
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Abstract

We have shown that affinity NMR can be used to edit a NMR spectrum so that ligands that have affinity to DNA can be observed in the presence of other nonbinding molecules. Diffusion encoded spectroscopy (DECODES) can be used to identify the binding ligands. We were able to identify Hoechst 33342 as binding to the Drew-Dickerson dodecamer d(CGCGAATTCGCG)2 in the presence of the nonbinding molecules adenine, adenosine, and thiamine. Affinity NMR appears to be readily applicable to DNA systems for the following reasons. (1) The relaxation rate of the DNA oligonucleotides is favorable, thus the signal intensity loss due to relaxation is not severe. (2) A comparison of the patterns of the DNA cross-peaks upon binding in the two-dimensional total correlation spectroscopy and correlation spectroscopy spectrum are easily performed, and the ligand signals in the two-dimensional DECODES spectrum can be readily identified. (3) The aromatic part of the DNA spectrum is devoid of 2D cross-peaks in these correlation spectra, greatly facilitating the interpretation of the bound ligand in the DECODES spectrum.

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AdenineAdenosineDNALigandsMagnetic Resonance SpectroscopyOligonucleotidesSpectrum AnalysisThiamineComparative StudyJournal Article

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R C Anderson, M Lin, M J Shapiro. 1999. Affinity NMR: decoding DNA binding.. https://doi.org/10.1021/cc980004o

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