A modified sensor chip for surface plasmon resonance enables a rapid determination of sequence specificity of DNA-binding proteins

FEBS Lett. 2003 Feb 11;536(1-3):151-6. doi: 10.1016/s0014-5793(03)00045-0.

Abstract

A novel method is described which rapidly determines specificity of DNA-binding proteins using a surface plasmon resonance (SPR) sensor chip. An oligohistidine-tagged DNA-binding domain of a transcription factor, NtERF2, was immobilised via nitrilotriacetic acid ligands to a sensor chip with an attenuated degree of carboxymethylation. DNA molecules were selected from a pool of randomised oligomers through binding to the immobilised protein and amplified by PCR. After several cycles of selection, during which binding was monitored by SPR, DNA sequences containing a consensus sequence were determined. The time necessary for one cycle is approximately 50 min, which is shorter than existing methods.

Publication types

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

MeSH terms

  • Base Sequence
  • Binding Sites
  • DNA / chemistry*
  • DNA / metabolism*
  • DNA-Binding Proteins / metabolism*
  • Kinetics
  • Nitrilotriacetic Acid / chemistry
  • Plant Proteins*
  • Surface Plasmon Resonance / methods*

Substances

  • DNA-Binding Proteins
  • Plant Proteins
  • ethylene-responsive element binding protein
  • DNA
  • Nitrilotriacetic Acid