Direct force measurement of single DNA–peptide interactions using atomic force microscopy |
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| Authors: | Ji W Chung Dongjin Shin June M Kwak Joonil Seog |
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| Institution: | 1. Fischell Department of Bioengineering, University of Maryland, , College Park, MD, 20742 USA;2. Institute of Antibody Research, Kangwon National University, , Chuncheon, Korea;3. Department of Cell Biology and Molecular Genetics, University of Maryland, , College Park, MD, 20742 USA;4. Department of Plant Molecular Systems Biotechnology and Crop Biotech Institute, Kyung Hee University, , Yongin, 446‐701 Korea;5. Department of Material Science and Engineering, University of Maryland, , College Park, MD, 20742 USA |
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| Abstract: | The selective interactions between DNA and miniature (39 residues) engineered peptide were directly measured at the single‐molecule level by using atomic force microscopy. This peptide (p007) contains an α‐helical recognition site similar to leucine zipper GCN4 and specifically recognizes the ATGAC sequence in the DNA with nanomolar affinity. The average rupture force was 42.1 pN, which is similar to the unbinding forces of the digoxigenin–antidigoxigenin complex, one of the strongest interactions in biological systems. The single linear fit of the rupture forces versus the logarithm of pulling rates showed a single energy barrier with a transition state located at 0.74 nm from the bound state. The smaller koff compared with that of other similar systems was presumably due to the increased stability of the helical structure by putative folding residues in p007. This strong sequence‐specific DNA–peptide interaction has a potential to be utilized to prepare well‐defined mechanically stable DNA–protein hybrid nanostructures. Copyright © 2013 John Wiley & Sons, Ltd. |
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| Keywords: | single‐molecule force spectroscopy p007 leucine zipper GCN4 dynamic force spectroscopy rupture force Bell– Evans model transition state |
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