Work

Nanopore Analysis of Single-Stranded Binding Protein Interactions with DNA

Year: 2015

Type: article

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.

Source: Langmuir

DOI: 10.1021/acs.langmuir.5b00457

Authors: Michael M. Marshall, Jan Ruzicka, Osama K. Zahid, Vincent C. Henrich, Ethan W. Taylor, Adam R. Hall

Institutions: University of North Carolina at Greensboro, Virginia Tech - Wake Forest University School of Biomedical Engineering & Sciences, Wake Forest University, Virginia Tech

Language: English

Topic: Nanopore and Nanochannel Transport Studies

Subfield: Biomedical Engineering

Field: Engineering

Domain: Physical Sciences

Open Access status: green

Funder: North Carolina Biotechnology Center

Award: G4412692325