Two-Step FRET as a Structural Tool

Watrob, Heather M.; Pan, Chia Pin; Barkley, Mary D.

doi:10.1021/ja034564p

Cited by 146 publications

(146 citation statements)

References 43 publications

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“…Single Molecule Three-color FRET Setup-We employed a one-donor/two-acceptor scheme (37,38) with Cy3 (donor), Atto647N (acceptor 1), and Cy5.5 (acceptor 2). Fluorescence intensities from the three fluorophores (Cy3, Atto647N, and Cy5.5) were spectrally separated into three regions by dichroic mirrors (Chroma Technology, Bellows Falls, VT) (see Fig.…”

Section: Methodsmentioning

confidence: 99%

Dynamics of Nucleosome Assembly and Effects of DNA Methylation

Lee

Yue

et al. 2015

Journal of Biological Chemistry

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Section: Methodsmentioning

confidence: 99%

Dynamics of Nucleosome Assembly and Effects of DNA Methylation

Lee

Yue

et al. 2015

Journal of Biological Chemistry

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“…For donors and acceptors that randomize by rotational diffusion prior to energy transfer, the magnitude of k 2 is assumed to be 2/3. 25,31 The refractive index (n) of the medium is 1.4, and the quantum yield (Q D ) of the donor in the absence of an acceptor is measured to be 3.8 Â 10…”

Section: Spectroscopic Studiesmentioning

confidence: 99%

Photoselective excited state dynamics in ZnO–Au nanocomposites and their implications in photocatalysis and dye-sensitized solar cells

Sarkar

Makhal

Bora

et al. 2011

Phys. Chem. Chem. Phys.

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Improving the performance of photoactive solid-state devices begins with systematic studies of the metal-semiconductor nanocomposites (NCs) upon which such devices are based. Here, we report the photo-dependent excitonic mechanism and the charge migration kinetics in a colloidal ZnO-Au NC system. By using a picosecond-resolved Fo¨rster resonance energy transfer (FRET) technique, we have demonstrated that excited ZnO nanoparticles (NPs) resonantly transfer visible optical radiation to the Au NPs, and the quenching of defect-mediated visible emission depends solely on the excitation level of the semiconductor. The role of the gold layer in promoting photolytic charge transfer, the activity of which is dependent upon the degree of excitation, was probed using methylene blue (MB) reduction at the semiconductor interface. Incident photon-tocurrent efficiency measurements show improved charge injection from a sensitizing dye to a semiconductor electrode in the presence of gold in the visible region. Furthermore, the short-circuit current density and the energy conversion efficiency of the ZnO-Au NP based dye-sensitized solar cell (DSSC) are much higher than those of a DSSC comprised of only ZnO NP. Our results represent a new paradigm for understanding the mechanism of defect-state passivation and photolytic activity of the metal component in metal-semiconductor nanocomposite systems.

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“…have been used extensively to determine numerous proteinprotein interactions (26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37). These studies include positioning troponin residues on the actin filament (28), determining the global structure of the VS ribozyme (29), mapping the antagonist-binding site of serotonin type 3 receptor (30), determining the structural organization of the synaptic exocytosis core (31), and defining the orientation of MRE-binding transcription factor 1 (32), as well as analyzing the spatial relationships between proteins of the nuclear pore complex (33).…”

mentioning

confidence: 99%

A fluorescence resonance energy transfer-derived structure of a quantum dot-protein bioconjugate nanoassembly

Medintz

Konnert²,

Clapp³

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

272

291

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The first generation of luminescent semiconductor quantum dot (QD)-based hybrid inorganic biomaterials and sensors is now being developed. It is crucial to understand how bioreceptors, especially proteins, interact with these inorganic nanomaterials. As a model system for study, we use Rhodamine red-labeled engineered variants of Escherichia coli maltose-binding protein (MBP) coordinated to the surface of 555-nm emitting CdSe-ZnS core-shell QDs. Fluorescence resonance energy transfer studies were performed to determine the distance from each of six unique MBP-Rhodamine red dye-acceptor locations to the center of the energy-donating QD. In a strategy analogous to a nanoscale global positioning system determination, we use the intraassembly distances determined from the fluorescence resonance energy transfer measurements, the MBP crystallographic coordinates, and a least-squares approach to determine the orientation of the MBP relative to the QD surface. Results indicate that MBP has a preferred orientation on the QD surface. The refined model is in agreement with other evidence, which indicates coordination of the protein to the QD occurs by means of its C-terminal pentahistidine tail, and the size of the QD estimated from the model is in good agreement with physical measurements of QD size. The approach detailed here may be useful in determining the orientation of proteins in other hybrid protein-nanoparticle materials. To our knowledge, this is the first structural model of a hybrid luminescent QD-protein receptor assembly elucidated by using spectroscopic measurements in conjunction with crystallographic and other data.maltose-binding protein ͉ three-dimensional structure ͉ nanotechnology ͉ nanocrystal T he burgeoning field of nanotechnology promises to revolutionize many scientific fields, and the first generation of functional hybrid nanomaterials exploring the interface between biology and materials science is now being developed and prototyped (1-3). One exciting avenue of biomaterials research involves protein-nanomaterial composites (2-4). Proteins lend many of their unique properties to these hybrid materials, such as: assisting in ordered self-assembly processes such as that of Pd nanoparticles assembled on tubulin or viral assembly of orientated nanowires (5, 6), engendering exquisite biorecognition properties such as the receptors used in hybrid nanocrystal biosensors (7), and catalyzing useful electrochemical and cleavage reactions (2, 8). Of critical importance in developing these materials is a fundamental understanding of how proteins or bioreceptors interact with inorganic nanomaterials.The unique properties of luminescent colloidal semiconductor nanocrystals or quantum dots (QDs) have recently been incorporated into hybrid functional nanoassemblies. Cadmium selenide-zinc sulfide (CdSe-ZnS) core-shell QDs, in particular, have exceptional photochemical stability and relatively high quantum yields, as well as broad excitation and size-tunable photoluminescence spectra with narrow emission bandwidt...

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Two-Step FRET as a Structural Tool

Cited by 146 publications

References 43 publications

Dynamics of Nucleosome Assembly and Effects of DNA Methylation

Dynamics of Nucleosome Assembly and Effects of DNA Methylation

Photoselective excited state dynamics in ZnO–Au nanocomposites and their implications in photocatalysis and dye-sensitized solar cells

A fluorescence resonance energy transfer-derived structure of a quantum dot-protein bioconjugate nanoassembly

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