Nano-mechanical measurements of protein-DNA interactions with a silicon nitride pulley.
Proteins adhere to DNA at locations and with strengths that depend on the protein conformation, the underlying DNA sequence and the ionic content of the solution. A facile technique to probe the positions and strengths of protein-DNA binding would aid in understanding these important interactions. Here, we describe a 'DNA pulley' for position-resolved nano-mechanical measurements of protein-DNA interactions. A molecule of λ DNA is tethered by one end to a glass surface, and by the other end to a magnetic bead. The DNA is stretched horizontally by a magnet, and a nanoscale knife made of silicon nitride is manipulated to contact, bend and scan along the DNA. The mechanical profile of the DNA at the contact with the knife is probed via nanometer-precision optical tracking of the magnetic bead. This system enables detection of protein bumps on the DNA and localization of their binding sites. We study theoretically the technical requirements to detect mechanical heterogeneities in the DNA itself.
Department of Chemistry and Chemical Biology and Department of Physics, Harvard University and Howard Hughes Medical Institute, Cambridge, MA 02138, USA.
Min Ju
MJ
Shon
Department of Chemistry and Chemical Biology and Department of Physics, Harvard University and Howard Hughes Medical Institute, Cambridge, MA 02138, USA cohen@chemistry.harvard.edu.
Adam E
AE
Cohen
D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D036103:Nanotechnology / Q000379:methods*; D011485:Protein Binding; D017655:Silicon Compounds / Q000737:chemistry*
D004268:DNA-Binding Proteins; D017655:Silicon Compounds; D004247:DNA; C032734:silicon nitride