ArticleProceedings of the National Academy of Sciences of the United States of America2004
Measuring single-molecule nucleic acid dynamics in solution by two-color filtered ratiometric fluorescence correlation spectroscopy.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed.
- Evidence of a Three-State Mechanism in DNA Hairpin Folding.Journal of the American Chemical Society · 2025Article
- Stacking-induced fluorescence increase reveals allosteric interactions through DNA.Nucleic acids research · 2018Article
- Gene editing in the context of an increasingly complex genome.BMC genomics · 2018Review
- Förster resonance energy transfer: Role of diffusion of fluorophore orientation and separation in observed shifts of FRET efficiency.PloS one · 2017Article
- Article
- Different fluorophore labeling strategies and designs affect millisecond kinetics of DNA hairpins.Molecules (Basel, Switzerland) · 2014Article
- Roles of long-range tertiary interactions in limiting dynamics of the Tetrahymena group I ribozyme.Journal of the American Chemical Society · 2014Article
- Imaging the cell surface and its organization down to the level of single molecules.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2013Review
- Label-free single-molecule detection of DNA-hybridization kinetics with a carbon nanotube field-effect transistor.Nature nanotechnology · 2011Article
- FRET fluctuation spectroscopy of diffusing biopolymers: contributions of conformational dynamics and translational diffusion.The journal of physical chemistry. B · 2010Article
- Single-molecule biophysics: at the interface of biology, physics and chemistry.Journal of the Royal Society, Interface · 2008Review
- RNA folding: conformational statistics, folding kinetics, and ion electrostatics.Annual review of biophysics · 2008Review
- Fluorescence from diffusing single molecules illuminates biomolecular structure and dynamics.Journal of fluorescence · 2007Review
- Dynamics of equilibrium structural fluctuations of apomyoglobin measured by fluorescence correlation spectroscopy.Proceedings of the National Academy of Sciences of the United States of America · 2007Article
- Dynamics of synaptic SfiI-DNA complex: single-molecule fluorescence analysis.Biophysical journal · 2007Article
- Photobleaching pathways in single-molecule FRET experiments.Journal of the American Chemical Society · 2007Article
- Molecular fluorescence, phosphorescence, and chemiluminescence spectrometry.Analytical chemistry · 2006Article
- Single-molecule fluorescence studies of protein folding and conformational dynamics.Chemical reviews · 2006Review
- The initial step of DNA hairpin folding: a kinetic analysis using fluorescence correlation spectroscopy.Nucleic acids research · 2006Article
- Single-molecule spectroscopic determination of lac repressor-DNA loop conformation.Biophysical journal · 2005Article
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5 authors.
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No grant is acknowledged in the PubMed record.
Abstract
This work presents a general method for determining single-molecule intramolecular dynamics in biomolecules by using a reporter fluorophore, whose fluorescence is quenched or partially quenched as a result of intramolecular motion, and a remote observer fluorophore. These fluorophores were excited independently with two different lasers, and the ratio of the two fluorophores' fluorescence was calculated. The time-varying ratio was then filtered to reduce contributions from molecules outside the overlapped laser volume and then correlated. The rates of opening and closing of a DNA hairpin were measured by using both fluorescence correlation spectroscopy and this method for comparison. We found at 50 pM, where molecules were studied one by one as they diffused through the probe volume, we obtained accurate opening and closing rates and could also measure dynamic heterogeneity. To demonstrate applicability to a more complex biological molecule we then probed intramolecular motions in the dimer of a human telomerase RNA fragment (hTR(380-444)), in the presence of an excess of monomer. The motion was found to occur on the time scale of 180-750 micros and slowed with increasing magnesium ion concentration. Blocking experiments using complementary oligonucleotides suggested that the motion involves substantial changes in dimer tertiary structure. This method appears to be a general method for selectively studying intramolecular motion in large biomolecules or complexes.
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