Computational Design of Transmembrane Pores

By Chunfu Xu, P. Lu, T. M. G. El-Din, X. Pei, Matthew C. Johnson, A. Uyeda, M. J. Bick, Qi Xu, D. Jiang, H. Bai and 15 more. 2020

141 citations recorded by the source corpus.

Transmembrane channels and pores have key roles in fundamental biological processes1 and in biotechnological applications such as DNA nanopore sequencing2–4, resulting in considerable interest in the design of pore-containing proteins. Synthetic amphiphilic peptides have been found to form ion channels5,6, and there have been recent advances in de novo membrane protein design7,8 and in redesigning naturally occurring channel-containing proteins9,10. However, the de novo design of stable, well-defined transmembrane protein pores that are capable of conducting ions selectively or are large enough to enable the passage of small-molecule fluorophores remains an outstanding challenge11,12. Here we report the computational design of protein pores formed by two concentric rings of α-helices that are stable and monodisperse in both their water-soluble and their transmembrane forms. Crystal structures of the water-soluble forms of a 12-helical pore and a 16-helical pore closely match the computational design models. Patch-clamp electrophysiology experiments show that, when expressed in insect cells, the transmembrane form of the 12-helix pore enables the passage of ions across the membrane with high selectivity for potassium over sodium; ion passage is blocked by specific chemical modification at the pore entrance. When incorporated into liposomes using in vitro protein synthesis, the transmembrane form of the 16-helix pore—but not the 12-helix pore—enables the passage of biotinylated Alexa Fluor 488. A cryo-electron microscopy structure of the 16-helix transmembrane pore closely matches the design model. The ability to produce structurally and functionally well-defined transmembrane pores opens the door to the creation of designer channels and pores for a wide variety of applications. An approach for the design of protein pores is demonstrated by the computational design and subsequent experimental expression of both an ion-selective and a large transmembrane pore.

Subfields: Biomedical Engineering, Molecular Biology

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In Biomedical Engineering

  1. Tanja Kortemme
  2. Xingjie Pan

In Molecular Biology

  1. Xingjie Pan
  2. Tanja Kortemme
  3. David Baker
  4. Jeffrey J. Gray
  5. I. Samish
  6. Colin A. Smith
  7. Jens Meiler
  8. Noah Ollikainen

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