Study of polynucleotide conformation by resolution‐enhanced ultraviolet spectroscopy

Self‐deconvolution and the fourth derivative of ultraviolet absorption spectra have been used to study stacked single‐stranded and double‐helix structures of different cytosine‐containing polynucleotides for the first time. These compounds were studied under different solution conditions (pH and organic solvents) and at low temperatures. The red shift of the lower band (B 2u band plus possibly some n →π* transition) of the absorption spectra in the cytosine‐containing polynucleotides and the appearance of new peaks in the deconvoluted and derivative spectra in the 280–310 nm region are attributed mainly to cytosine‐cytosine stacking interactions. In particular, the fourth‐derivative peaks at wavelengths higher than 290 nm can be associated to coupling of electronic transitions of cytosine bases. The nature of the electronic transitions producing the absorption bands which are resolved in the aforementioned fourth‐derivative peaks is discussed. It is concluded that the resolution‐enhancement techniques used in this work, i.e. self‐deconvolution and fourth derivative, complement each other and are useful methods to study structural changes of single‐stranded and double‐stranded polynucleotides allowing, at the same time, more information to be obtained about specific stacking interactions than classical absorption spectrophotometry.