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An engineered dimeric protein pore that spans adjacent lipid bilayers

Author

Listed:
  • Shiksha Mantri

    (University of Oxford)

  • K. Tanuj Sapra

    (University of Oxford)

  • Stephen Cheley

    (University of Oxford
    Present address: Department of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, Canada T6G 2E1)

  • Thomas H. Sharp

    (University of Oxford)

  • Hagan Bayley

    (University of Oxford)

Abstract

The bottom-up construction of artificial tissues is an underexplored area of synthetic biology. An important challenge is communication between constituent compartments of the engineered tissue, and between the engineered tissue and additional compartments, including extracellular fluids, further engineered tissue and living cells. Here we present a dimeric transmembrane pore that can span two adjacent lipid bilayers, and thereby allow aqueous compartments to communicate. Two heptameric staphylococcal α-hemolysin pores were covalently linked in an aligned cap-to-cap orientation. The structure of the dimer, (α7)2, was confirmed by biochemical analysis, transmission electron microscopy and single-channel electrical recording. We show that one of two β-barrels of (α7)2 can insert into the lipid bilayer of a small unilamellar vesicle, while the other spans a planar lipid bilayer. The (α7)2 pores spanning two bilayers were also observed by transmission electron microscopy.

Suggested Citation

  • Shiksha Mantri & K. Tanuj Sapra & Stephen Cheley & Thomas H. Sharp & Hagan Bayley, 2013. "An engineered dimeric protein pore that spans adjacent lipid bilayers," Nature Communications, Nature, vol. 4(1), pages 1-10, June.
    • Handle: RePEc:nat:natcom:v:4:y:2013:i:1:d:10.1038_ncomms2726
    DOI: 10.1038/ncomms2726
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    Cited by:

    1. Alexander Harjung & Alessandro Fracassi & Neal K. Devaraj, 2024. "Encoding extracellular modification of artificial cell membranes using engineered self-translocating proteins," Nature Communications, Nature, vol. 15(1), pages 1-11, December.
    2. Swarup Dey & Adam Dorey & Leeza Abraham & Yongzheng Xing & Irene Zhang & Fei Zhang & Stefan Howorka & Hao Yan, 2022. "A reversibly gated protein-transporting membrane channel made of DNA," Nature Communications, Nature, vol. 13(1), pages 1-12, December.

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