FRET fluctuation spectroscopy: exploring the conformational dynamics of a DNA hairpin loop

MI Wallace, L Ying, S Balasubramanian… - The Journal of …, 2000 - ACS Publications
MI Wallace, L Ying, S Balasubramanian, D Klenerman
The Journal of Physical Chemistry B, 2000ACS Publications
The motions of a dye-labeled DNA hairpin loop (Cy5-5 '-GGGTT-(A) 30-AACCC-3 '-TMR)
have been investigated through the fluctuations in proximity ratio from fluorescence
resonance energy transfer (FRET). We examine three solution conditions:(1) MilliQ water,(2)
Tris-EDTA buffer, and (3) Tris-EDTA buffer plus an excess of DNA complementary to the
loop sequence,(T) 30. Correlations in proximity ratio show submillisecond dynamics. Static
heterogeneity is revealed from the distribution of proximity ratio amplitudes. The observed …
The motions of a dye-labeled DNA hairpin loop (Cy5-5‘-GGGTT-(A)30-AACCC-3‘-TMR) have been investigated through the fluctuations in proximity ratio from fluorescence resonance energy transfer (FRET). We examine three solution conditions:  (1) MilliQ water, (2) Tris-EDTA buffer, and (3) Tris-EDTA buffer plus an excess of DNA complementary to the loop sequence, (T)30. Correlations in proximity ratio show submillisecond dynamics. Static heterogeneity is revealed from the distribution of proximity ratio amplitudes. The observed stretched exponential kinetics are consistent with a model based on the transition between two states over a complex energy landscape.
ACS Publications
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