Platform for in situ real-time measurement of protein-induced conformational changes of DNA

Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1397-401. doi: 10.1073/pnas.0912182107. Epub 2010 Jan 4.

Abstract

A platform for in situ and real-time measurement of protein-induced conformational changes in dsDNA is presented. We combine electrical orientation of surface-bound dsDNA probes with an optical technique to measure the kinetics of DNA conformational changes. The sequence-specific Escherichia coli integration host factor is utilized to demonstrate protein-induced bending upon binding of integration host factor to dsDNA probes. The effects of probe surface density on binding/bending kinetics are investigated. The platform can accommodate individual spots of microarrayed dsDNA on individually controlled, lithographically designed electrodes, making it amenable for use as a high throughput assay.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • DNA Probes / analysis*
  • DNA Probes / chemistry
  • DNA, Bacterial / analysis*
  • DNA, Bacterial / chemistry
  • Electrodes
  • Escherichia coli / chemistry*
  • Escherichia coli Proteins / analysis*
  • Escherichia coli Proteins / chemistry
  • Gold / chemistry
  • Nucleic Acid Conformation*
  • Spectrometry, Fluorescence
  • Time Factors

Substances

  • DNA Probes
  • DNA, Bacterial
  • Escherichia coli Proteins
  • Gold