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1H/31P NMR techniques were applied to analyze the binding mode of guanosine 2'-monophosphate (2'-GMP) to histone. To date, no structures of the complex comprising 2'-GMP and histone have been deposited in Protein Data Bank. Because the 31P nucleus can be a selective marker of phosphorylated compounds, the combined use of 1H and 31P NMR spectroscopy has been applied to investigate these molecular interactions. The complex formation was initially confirmed by 31P-diffusion ordered spectroscopy and 31P-T1 measurements. In 1H1H saturation tra