Capture of unstable protein complex on the streptavidin-coated single-walled carbon nanotubes
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Purification of unstable protein complexes is a bottleneck for investigation of their 3D structure and in protein–protein interaction studies. In this paper, we demonstrate that streptavidin-coated single-walled carbon nanotubes (Strep•SWNT) can be used to capture the biotinylated DNA–EcoRI complexes on a 2D surface and in solution using atomic force microscopy and electrophoresis analysis, respectively. The restriction enzyme EcoRI forms unstable complexes with DNA in the absence of Mg2+. Capturing the EcoRI–DNA complexes on the Strep•SWNT succeeded in the absence of Mg2+, demonstrating that the Strep•SWNT can be used for purifying unstable protein complexes.
KeywordsUnstable protein complex Carbon nanotube Streptavidin EcoRI
The authors gratefully acknowledge the financial support from The Netherlands Organisation for Scientific Research (NWO), Veni Grant 2009 (Project Number: 700.59.407), Cyttron II: www.cyttryon.org, the National Natural Science Foundation of China (Grant No. 31200637), Jiangsu Basic Research Program (The Natural Science Fund No. SBK201240183), and the Science and Technology Support Program of Changzhou (Social Development Grant No. CE20125050). We thank René Olsthoorn, Claude Backendorf, and Hans den Dulk for technical support.
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