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

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 similar to50 min, which is shorter than existing methods. (C) 2003 Published by Elsevier Science B.V. on behalf of the Federation of European Biochemical Societies.