Pure spectra of actinomycin D bound to DNA at various ionic strengths extracted by the principal-component-analysis method.

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The interaction between actinomycin D (AMD) and native and heat-denatured DNA was studied spectrophotometrically at 25 °C. By applying the principal-component-analysis (PCA) method to the series of observed spectra, the number of light-absorbing components was determined. Two spectroscopically different species of bound AMD, i.e., AMD–DNA complexes, were formed when an aqueous AMD solution was titrated with either the native or the denatured DNA solution over a wide range of the nucleotide residue-to-AMD mixing ratios (P/D) between 0 and 30. The pure spectra of AMD bound to DNA were extracted in the P/D range 0–4 by the extended PCA method with an appropriate binding scheme. These bound-AMD spectra were affected by the DNA conformation but not by the addition of NaCl (1–500 mmol dm−3). The fractions of bound AMD were calculated as a function of P/D.
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