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A streptavidin variant with slower biotin dissociation and increased mechanostability

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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Figure 1: Traptavidin has a slower dissociation rate from biotin conjugates compared to streptavidin.
Figure 2: Traptavidin has increased thermostability compared to streptavidin.
Figure 3: Traptavidin has increased mechanical stability compared to streptavidin.

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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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Contributions

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.

Corresponding author

Correspondence to Mark Howarth.

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Competing interests

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 Text and Figures

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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