期刊
BIOSENSORS & BIOELECTRONICS
卷 21, 期 8, 页码 1599-1605出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2005.07.018
关键词
transcription factors; SP1; NF-kappa B; cantilever sensor; proteomics; DNA binding protein
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-kappa B, 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-kappa B. 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. (c) 2005 Elsevier B.V. All rights reserved.
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