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Nano Lett. 2008 Jun;8(6):1631-7. doi: 10.1021/nl080316l. Epub 2008 Apr 30.

Functional characterization and atomic force microscopy of a DNA repair protein conjugated to a quantum dot.

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1
Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, NIH, Research Triangle Park, NC 27709, USA.

Abstract

Quantum dots (QDs) possess highly desirable optical properties that make them ideal fluorescent labels for studying the dynamic behavior of proteins. However, a lack of characterization methods for reliably determining protein-quantum dot conjugate stoichiometry and functionality has impeded their widespread use in single-molecule studies. We used atomic force microscopic (AFM) imaging to demonstrate the 1:1 formation of UvrB-QD conjugates based on an antibody-sandwich method. We show that an agarose gel-based electrophoresis mobility shift assay and AFM can be used to evaluate the DNA binding function of UvrB-QD conjugates. Importantly, we demonstrate that quantum dots can serve as a molecular marker to unambiguously identify the presence of a labeled protein in AFM images.

PMID:
18444686
PMCID:
PMC3941028
DOI:
10.1021/nl080316l
[Indexed for MEDLINE]
Free PMC Article
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