Details

Autor(en) / Beteiligte

• Xu, Chunfu
• Lu, Peilong
• Gamal El-Din, Tamer M.
• Pei, Xue Y.
• Johnson, Matthew C.
• Uyeda, Atsuko
• Bick, Matthew J.
• Xu, Qi
• Jiang, Daohua
• Bai, Hua
• Reggiano, Gabriella
• Hsia, Yang
• Brunette, T J
• Dou, Jiayi
• Ma, Dan
• Lynch, Eric M.
• Boyken, Scott E.
• Huang, Po-Ssu
• Stewart, Lance
• DiMaio, Frank
• Kollman, Justin M.
• Luisi, Ben F.
• Matsuura, Tomoaki
• Catterall, William A.
• Baker, David

Titel

Computational design of transmembrane pores

Ist Teil von

• Nature (London), 2020-09, Vol.585 (7823), p.129-134

Ort / Verlag

London: Nature Publishing Group UK

Erscheinungsjahr

2020

Quelle

MEDLINE

Beschreibungen/Notizen

• Transmembrane channels and pores have key roles in fundamental biological processes 1 and in biotechnological applications such as DNA nanopore sequencing 2 – 4 , resulting in considerable interest in the design of pore-containing proteins. Synthetic amphiphilic peptides have been found to form ion channels 5 , 6 , and there have been recent advances in de novo membrane protein design 7 , 8 and in redesigning naturally occurring channel-containing proteins 9 , 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 challenge 11 , 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.