Fluorescence Resonance Energy Transfer Between Quantum Dot Donors and Dye-Labeled Protein Acceptors

We used luminescent CdSe−ZnS core−shell quantum dots (QDs) as energy donors in fluorescent resonance energy transfer (FRET) assays. Engineered maltose binding protein (MBP) appended with an oligohistidine tail and labeled with an acceptor dye (Cy3) was immobilized on the nanocrystals via a noncovale...

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Bibliographic Details
Published in:Journal of the American Chemical Society 2004-01, Vol.126 (1), p.301-310
Main Authors: Clapp, Aaron R, Medintz, Igor L, Mauro, J. Matthew, Fisher, Brent R, Bawendi, Moungi G, Mattoussi, Hedi
Format: Article
Language:English
Subjects:
Cadmium Compounds - chemistry
Carbocyanines - chemistry
Carrier Proteins - chemistry
Condensed matter: electronic structure, electrical, magnetic, and optical properties
Exact sciences and technology
Fluorescence Resonance Energy Transfer - methods
Fluorescent Dyes - chemistry
Histidine - chemistry
Luminescent Measurements
Maltose-Binding Proteins
Optical properties and condensed-matter spectroscopy and other interactions of matter with particles and radiation
Optical properties of low-dimensional, mesoscopic, and nanoscale materials and structures
Physics
Quantum Dots
Selenium Compounds - chemistry
Spectrometry, Fluorescence
Sulfides - chemistry
Titrimetry
Zinc Compounds - chemistry
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Summary:We used luminescent CdSe−ZnS core−shell quantum dots (QDs) as energy donors in fluorescent resonance energy transfer (FRET) assays. Engineered maltose binding protein (MBP) appended with an oligohistidine tail and labeled with an acceptor dye (Cy3) was immobilized on the nanocrystals via a noncovalent self-assembly scheme. This configuration allowed accurate control of the donor−acceptor separation distance to a range smaller than 100 Å and provided a good model system to explore FRET phenomena in QD−protein−dye conjugates. This QD−MBP conjugate presents two advantages:  (1) it permits one to tune the degree of spectral overlap between donor and acceptor and (2) provides a unique configuration where a single donor can interact with several acceptors simultaneously. The FRET signal was measured for these complexes as a function of both degree of spectral overlap and fraction of dye-labeled proteins in the QD conjugate. Data showed that substantial acceptor signals were measured upon conjugate formation, indicating efficient nonradiative exciton transfer between QD donors and dye-labeled protein acceptors. FRET efficiency can be controlled either by tuning the QD photoemission or by adjusting the number of dye-labeled proteins immobilized on the QD center. Results showed a clear dependence of the efficiency on the spectral overlap between the QD donor and dye acceptor. Apparent donor−acceptor distances were determined from efficiency measurements and corresponding Förster distances, and these results agreed with QD bioconjugate dimensions extracted from structural data and core size variations among QD populations.
ISSN:0002-7863
1520-5126
DOI:10.1021/ja037088b