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Engineered nanostructured β-sheet peptides protect membrane proteins

Nature Methods volume 10, pages 759761 (2013) | Download Citation

Abstract

We designed β-strand peptides that stabilize integral membrane proteins (IMPs). β-strand peptides self-assemble in solution as filaments and become restructured upon association with IMPs; resulting IMP–β-strand peptide complexes resisted aggregation when diluted in detergent-free buffer and were visible as stable, single particles with low detergent background in electron micrographs. β-strand peptides enabled clear visualization of flexible conformations in the highly dynamic ATP-binding cassette (ABC) transporter MsbA.

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Acknowledgements

This work was supported by grants from US National Institute of General Medical Sciences (P50 GM073197 and R01 GM095573). We thank J. Kelly for use of his circular dichroism instrument; F. Romesberg for use of his Fourier transfer infrared spectrometer; G. Chang (University of California, San Diego), S. Choe (Salk Institute for Biological Studies) and S. Sligar (University of Illinois Urbana-Champaign) for providing recombinant DNA encoding MsbA, KcsA and MSP1D1, respectively; J. Kelly, G. Chang, A. Ward, V. Cherezov, P. Dawson and W. Chen for discussions and comments; and E. Hildebrandt for editorial assistance.

Author information

Author notes

    • Rituparna Sinha Roy

    Present address: Department of Biological Sciences, Department of Chemical Sciences, Indian Institute of Science Education and Research, Kolkata, India.

    • Houchao Tao
    • , Sung Chang Lee
    •  & Arne Moeller

    These authors contributed equally to this work.

Affiliations

  1. Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, California, USA.

    • Houchao Tao
    • , Sung Chang Lee
    • , Arne Moeller
    • , Rituparna Sinha Roy
    • , Fai Yiu Siu
    • , Raymond C Stevens
    • , Clinton S Potter
    • , Bridget Carragher
    •  & Qinghai Zhang
  2. The National Resource for Automated Molecular Microscopy, The Scripps Research Institute, La Jolla, California, USA.

    • Arne Moeller
    • , Clinton S Potter
    •  & Bridget Carragher
  3. Department of Chemistry, The Scripps Research Institute, La Jolla, California, USA.

    • Jörg Zimmermann

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Contributions

Peptides were synthesized by H.T. and R.S.R.; peptide assembly was characterized by H.T., S.C.L., A.M. and J.Z.; protein stabilizations were performed by H.T. and S.C.L.; GLR was prepared by F.Y.S. with oversight by R.C.S.; MsbA samples for electron microscopy were prepared by S.C.L.; the electron microscopy study was performed by A.M. with oversight by B.C. and C.S.P.; and data were interpreted by all authors. The manuscript was written by H.T., S.C.L, A.M., B.C. and Q.Z.; and the study was conceived and overseen by Q.Z.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Qinghai Zhang.

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DOI

https://doi.org/10.1038/nmeth.2533

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