title: Nanopore Analysis of Single-Stranded Binding Protein Interactions with DNA.
abstract: We study the binding of E. coli single-stranded binding protein (SSB) to single-stranded DNA (ssDNA) using a solid-state nanopore assay. We find that saturated nucleoprotein complexes can be distinguished easily from free SSB, ssDNA, or double-stranded DNA individually and demonstrate that the high affinity of SSB for ssDNA can be exploited to achieve high-fidelity differentiation from duplex molecules in a mixture. We then study nucleoprotein filament formation by systematically varying the amount of SSB relative to ssDNA. We observe a concomitant shift in the mean amplitude of electrical events that is consistent with weakly cooperative binding. Finally, we compare circular and linearized ssDNA saturated with SSB and use the results to infer structural details of the nucleoprotein complex.
authors.affiliation: †Joint School of Nanoscience and Nanoengineering and ‡Center for Biotechnology, Genomics, and Health Research, University of North Carolina at Greensboro, Greensboro, North Carolina 27401, United States.|§Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences and ∥Comprehensive Cancer Center, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27101, United States.
authors.forename: Michael M
authors.initials: MM
authors.lastname: Marshall
authors.affiliation: †Joint School of Nanoscience and Nanoengineering and ‡Center for Biotechnology, Genomics, and Health Research, University of North Carolina at Greensboro, Greensboro, North Carolina 27401, United States.|§Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences and ∥Comprehensive Cancer Center, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27101, United States.
authors.forename: Jan
authors.initials: J
authors.lastname: Ruzicka
authors.affiliation: †Joint School of Nanoscience and Nanoengineering and ‡Center for Biotechnology, Genomics, and Health Research, University of North Carolina at Greensboro, Greensboro, North Carolina 27401, United States.|§Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences and ∥Comprehensive Cancer Center, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27101, United States.
authors.forename: Osama K
authors.initials: OK
authors.lastname: Zahid
authors.affiliation: †Joint School of Nanoscience and Nanoengineering and ‡Center for Biotechnology, Genomics, and Health Research, University of North Carolina at Greensboro, Greensboro, North Carolina 27401, United States.|§Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences and ∥Comprehensive Cancer Center, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27101, United States.
authors.forename: Vincent C
authors.initials: VC
authors.lastname: Henrich
authors.affiliation: †Joint School of Nanoscience and Nanoengineering and ‡Center for Biotechnology, Genomics, and Health Research, University of North Carolina at Greensboro, Greensboro, North Carolina 27401, United States.|§Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences and ∥Comprehensive Cancer Center, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27101, United States.
authors.forename: Ethan W
authors.initials: EW
authors.lastname: Taylor
authors.affiliation: †Joint School of Nanoscience and Nanoengineering and ‡Center for Biotechnology, Genomics, and Health Research, University of North Carolina at Greensboro, Greensboro, North Carolina 27401, United States.|§Virginia Tech-Wake Forest University School of Biomedical Engineering and Sciences and ∥Comprehensive Cancer Center, Wake Forest University School of Medicine, Winston-Salem, North Carolina 27101, United States.
authors.forename: Adam R
authors.initials: AR
authors.lastname: Hall
mesh_terms: D004277:DNA, Single-Stranded / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D055664:Electrochemical Techniques; D004926:Escherichia coli / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry*; D058608:Nanopores; D009698:Nucleoproteins / Q000737:chemistry*; D009994:Osmolar Concentration; D011485:Protein Binding
chemical_list: D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D009698:Nucleoproteins