Donor fluorescence decay analysis for energy transfer in double-helical DNA with various acceptor concentrations

Shin Ichi Murata, Józef Kuba, Grzegorz Piszczek, Ignacy Gryczynski, Joseph R. Lakowicz

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Abstract

We studied fluorescence resonance energy transfer between donors and acceptors bound to double-helical DNA. The donor Hoechst 33258 binds to the minor groove of DNA and the acceptor propidium iodide (PI) is an intercalator. The time-resolved donor decays were measured in the frequency domain. The donor decays were consistent with a random 1-dimensional distribution of acceptors. The decays were analyzed in terms of three 1-dimensional models: a random continuous acceptor distribution; acceptors placed on discrete lattice sites; and a cylindrical model with the acceptor in the center, and the donors on a cylinder surface. The data were well described by all three models. Interpretation in terms of continuous distribution of acceptors revealed a minimum donor to acceptor distance of 13 Å, which is 3 bp from the center of Hoechst 33252. These results suggest that PI is excluded from the 4 bp covered by Hoechst 33252 when it is bound to the minor groove of DNA. (C) 2000 John Wiley and Sons, Inc.

Original languageEnglish
Pages (from-to)306-315
Number of pages10
JournalBiopolymers - Biospectroscopy Section
Volume57
Issue number5
DOIs
Publication statusPublished - 29 Aug 2000

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Keywords

  • Acceptor
  • Donor
  • Double-helical DNA
  • Fluorescence resonance energy transfer

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