Article

A DNA tweezer-actuated enzyme nanoreactor

  • Nature Communications 4, Article number: 2127 (2013)
  • doi:10.1038/ncomms3127
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Abstract

The functions of regulatory enzymes are essential to modulating cellular pathways. Here we report a tweezer-like DNA nanodevice to actuate the activity of an enzyme/cofactor pair. A dehydrogenase and NAD+ cofactor are attached to different arms of the DNA tweezer structure and actuation of enzymatic function is achieved by switching the tweezers between open and closed states. The enzyme/cofactor pair is spatially separated in the open state with inhibited enzyme function, whereas in the closed state, enzyme is activated by the close proximity of the two molecules. The conformational state of the DNA tweezer is controlled by the addition of specific oligonucleotides that serve as the thermodynamic driver (fuel) to trigger the change. Using this approach, several cycles of externally controlled enzyme inhibition and activation are successfully demonstrated. This principle of responsive enzyme nanodevices may be used to regulate other types of enzymes and to introduce feedback or feed-forward control loops.

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Acknowledgements

This work is supported by an Army Research Office grant W911NF-11-1-0137 to H.Y. and Y.L., an Army Research Office MURI award W911NF-12-1-0420 to H.Y. and N.W., and a National Science Foundation grant 1033222 to N.W. and H.Y.; H.Y. and Y.L. is part of the Center for Bio-Inspired Solar Fuel Production, an Energy Frontier Research Center funded by the US Department of Energy, Office of Science, Office of Basic Energy Sciences under Award Number DE-SC0001016. H.Y. is supported by the Presidential Strategic Initiative Fund from Arizona State University.

Author information

Author notes

    • Minghui Liu
    •  & Jinglin Fu

    These authors contributed equally to this work

Affiliations

  1. Center for Single Molecule Biophysics, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA

    • Minghui Liu
    • , Jinglin Fu
    • , Yuhe Yang
    • , Yan Liu
    •  & Hao Yan
  2. Department of Chemistry and Biochemistry, Arizona State University, Tempe, Arizona 85287, USA

    • Minghui Liu
    • , Yuhe Yang
    • , Neal W. Woodbury
    • , Yan Liu
    •  & Hao Yan
  3. Center for Innovations in Medicine, Biodesign Institute, Arizona State University, Tempe, Arizona 85287, USA

    • Jinglin Fu
    •  & Neal W. Woodbury
  4. Department of Chemistry and iNANO, Centre for DNA Nanotechnology, Aarhus University, Aarhus C DK-8000, Denmark

    • Christian Hejesen
    •  & Kurt Gothelf

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Contributions

J.F. and H.Y. conceived the concepts. M.L. and J.F. designed and performed the experiments, and analyzed the data. C.H. developed the NAD–DNA conjugation chemistry. M.L., J.F. and Y.Y. optimized DNA assembly and protein/oligo conjugation. M.L. and J.F. wrote the manuscript. All the authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Jinglin Fu or Hao Yan.

Supplementary information

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  1. 1.

    Supplementary Information

    Supplementary Figures S1-S16, Supplementary Tables S1-S2, Supplementary Methods and Supplementary References

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