title: Functional characterization and atomic force microscopy of a DNA repair protein conjugated to a quantum dot.
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.
authors.affiliation: Laboratory of Molecular Genetics, National Institute of Environmental Health Sciences, NIH, Research Triangle Park, NC 27709, USA.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.forename: Ingrid
authors.initials: I
authors.lastname: Tessmer
authors.forename: Deborah L
authors.initials: DL
authors.lastname: Croteau
authors.forename: Dorothy A
authors.initials: DA
authors.lastname: Erie
authors.forename: Bennett
authors.initials: B
authors.lastname: Van Houten
mesh_terms: D001665:Binding Sites; D045643:DNA Repair Enzymes / Q000737:chemistry* / Q000648:ultrastructure*; D018625:Microscopy, Atomic Force / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D045663:Quantum Dots
chemical_list: D045643:DNA Repair Enzymes