Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

Single-pair fluorescence resonance energy transfer on freely diffusing molecules: observation of Förster distance dependence and subpopulations

Academic Article
uri icon
  • Overview
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Deniz, Ashok
  • Dahan, M.
  • Grunwell, J. R.
  • Ha, T. J.
  • Faulhaber, A. E.
  • Chemla, D. S.
  • Weiss, S.
  • Schultz, Peter

publication date

  • March 1999

journal

  • Proceedings of the National Academy of Sciences of the United States of America  Journal

abstract

  • Photon bursts from single diffusing donor-acceptor labeled macromolecules were used to measure intramolecular distances and identify subpopulations of freely diffusing macromolecules in a heterogeneous ensemble. By using DNA as a rigid spacer, a series of constructs with varying intramolecular donor-acceptor spacings were used to measure the mean and distribution width of fluorescence resonance energy transfer (FRET) efficiencies as a function of distance. The mean single-pair FRET efficiencies qualitatively follow the distance dependence predicted by Förster theory. Possible contributions to the widths of the FRET efficiency distributions are discussed, and potential applications in the study of biopolymer conformational dynamics are suggested. The ability to measure intramolecular (and intermolecular) distances for single molecules implies the ability to distinguish and monitor subpopulations of molecules in a mixture with different distances or conformational states. This is demonstrated by monitoring substrate and product subpopulations before and after a restriction endonuclease cleavage reaction. Distance measurements at single-molecule resolution also should facilitate the study of complex reactions such as biopolymer folding. To this end, the denaturation of a DNA hairpin was examined by using single-pair FRET.

subject areas

  • DNA
  • Energy Transfer
  • Nucleic Acid Conformation
  • Nucleic Acid Denaturation
  • Oligodeoxyribonucleotides
  • Photons
  • Spectrometry, Fluorescence
  • Urea
scroll to property group menus

Identity

PubMed Central ID

  • PMC22352

International Standard Serial Number (ISSN)

  • 0027-8424

Digital Object Identifier (DOI)

  • 10.1073/pnas.96.7.3670

PubMed ID

  • 10097095
scroll to property group menus

Additional Document Info

start page

  • 3670

end page

  • 3675

volume

  • 96

issue

  • 7

©2021 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support