Abstract
Streptavidin binds biotin conjugates with exceptional stability but dissociation does occur, limiting its use in imaging, DNA amplification and nanotechnology. We identified a mutant streptavidin, traptavidin, with more than tenfold slower biotin dissociation, increased mechanical strength and improved thermostability; this resilience should enable diverse applications. FtsK, a motor protein important in chromosome segregation, rapidly displaced streptavidin from biotinylated DNA, whereas traptavidin resisted displacement, indicating the force generated by Ftsk translocation.
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Acknowledgements
Funding was provided by the Wellcome Trust (M.H., D.J.S. and E.C.), the Biotechnology and Biological Sciences Research Council (C.E.C.), the US National Institutes of Health (V.T.M. and C.C.), the National Science Foundation (V.T.M.), the University of Miami (V.T.M.) and Worcester College Oxford (M.H.). We thank J.E. Graham and I. Grainge for helpful discussion.
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C.E.C., M.H., E.C., V.T.M. and C.C. performed the research; M.H., V.T.M. and D.J.S. designed research; all analyzed the data; M.H., D.J.S. and C.E.C. wrote the manuscript.
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M.H. and C.E.C. are authors on a patent application regarding the streptavidin variant described in this study (UK patent application 0919102.4).
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Supplementary Figures 1–5, Supplementary Table 1 and Supplementary Note (PDF 6844 kb)
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Chivers, C., Crozat, E., Chu, C. et al. A streptavidin variant with slower biotin dissociation and increased mechanostability. Nat Methods 7, 391–393 (2010). https://doi.org/10.1038/nmeth.1450
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DOI: https://doi.org/10.1038/nmeth.1450
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