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Donor fluorescence decay analysis for energy transfer in double‐helical DNA with various acceptor concentrations
Author(s) -
Murata ShinIchi,
Kuśba Józef,
Piszczek Grzegorz,
Gryczynski Ignacy,
Lakowicz Joseph R.
Publication year - 2000
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/1097-0282(2000)57:5<306::aid-bip70>3.0.co;2-7
Subject(s) - chemistry , acceptor , fluorescence , förster resonance energy transfer , crystallography , dna , propidium iodide , minor groove , physics , apoptosis , biochemistry , programmed cell death , condensed matter physics , quantum mechanics
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. © 2000 John Wiley & Sons, Inc. Biopolymers (Biospectroscopy) 57: 306–315, 2000