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Multiple binding modes for Hoechst 33258 to DNA.

T Stokke, H B Steen

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society

|April 1, 1985

Summary

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The bisbenzimidazole Hoechst 33258 exhibits two distinct binding modes to DNA, influencing its fluorescence properties. High concentrations lead to DNA precipitation via cooperative interactions.

Area of Science:

Biochemistry
Molecular Biology
Biophysical Chemistry

Background:

Hoechst 33258 is a bisbenzimidazole dye commonly used to stain DNA.
Understanding dye-DNA interactions is crucial for molecular biology techniques.

Purpose of the Study:

To elucidate the binding modes of Hoechst 33258 to native DNA.
To characterize the fluorescence properties and precipitation behavior of the dye-DNA complex.

Main Methods:

Spectroscopic analysis of fluorescence emission and quantum yield.
Investigation of dye/phosphate ratios (D/P) to determine binding modes.
Assessment of precipitation kinetics and conditions.

Main Results:

Two binding modes were identified: Type 1 (low D/P) with high fluorescence quantum yield (emission at 460 nm) and Type 2 (0.05 < D/P < 0.4) with quenched fluorescence (emission around 490 nm).

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Precipitation of the dye-DNA complex occurred at D/P > 0.4, driven by cooperative interactions and strong electronic coupling between dye molecules.

Ethanol altered binding; Type 2 binding was inhibited, and DNA precipitation was prevented.

Conclusions:

Hoechst 33258 binding to DNA is concentration-dependent, leading to distinct fluorescence characteristics and precipitation.
The dye's interaction with DNA is influenced by solution conditions like ethanol presence.