Label free analysis of transcription factors using microcantilever arrays

Biosens Bioelectron. 2006 Feb 15;21(8):1599-605. doi: 10.1016/j.bios.2005.07.018. Epub 2005 Aug 30.

Abstract

We report the measurement of protein interaction with double-stranded DNA oligonucleotides using cantilever microarray technology. We investigated two different DNA-binding proteins, the transcription factors SP1 and NF-kappaB, using cantilever arrays as they allow label-free measurement of different biomolecular interactions in parallel. Double-stranded DNA oligonucleotides containing a specific binding site for a transcription factor were sensitized on gold-coated cantilevers. The binding of the transcription factor creates a surface stress, resulting in a bending of the cantilevers. Both transcription factors could be detected independently at concentrations of 80-100 nM. A concentration dependence of the bending signal was measured using concentrations from 100 to 400 nM of NF-kappaB. The experiments show that the recognition sequence of one transcription factor can serve as a reference for the other, highlighting the sequence specificity of transcription factor binding.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / analysis*
  • DNA / chemistry
  • DNA-Binding Proteins / analysis*
  • DNA-Binding Proteins / chemistry
  • Equipment Design
  • Equipment Failure Analysis
  • Protein Array Analysis / instrumentation*
  • Protein Array Analysis / methods
  • Refractometry / instrumentation*
  • Refractometry / methods
  • Staining and Labeling
  • Transcription Factors / analysis*
  • Transcription Factors / chemistry

Substances

  • DNA-Binding Proteins
  • Transcription Factors
  • DNA