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Computational design of transmembrane pores.

Abstract：

: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.

journal_name

Nature

journal_title

Nature

authors

Xu C,Lu P,Gamal El-Din TM,Pei XY,Johnson MC,Uyeda A,Bick MJ,Xu Q,Jiang D,Bai H,Reggiano G,Hsia Y,Brunette TJ,Dou J,Ma D,Lynch EM,Boyken SE,Huang PS,Stewart L,DiMaio F,Kollman JM,Luisi BF,Matsuura T,Catterall

doi

10.1038/s41586-020-2646-5

subject

Has Abstract

pub_date

2020-09-01 00:00:00

pages

129-134

issue

7823

eissn

0028-0836

issn

1476-4687

pii

10.1038/s41586-020-2646-5

journal_volume

585

pub_type

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Computational design of transmembrane pores.