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-Structure paper
タイトル | Computational design of transmembrane pores. |
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ジャーナル・号・ページ | Nature, Vol. 585, Issue 7823, Page 129-134, Year 2020 |
掲載日 | 2020年8月26日 |
著者 | Chunfu Xu / Peilong Lu / Tamer M Gamal El-Din / Xue Y Pei / Matthew C Johnson / Atsuko Uyeda / Matthew J Bick / Qi Xu / Daohua Jiang / Hua Bai / Gabriella Reggiano / Yang Hsia / T J Brunette / Jiayi Dou / Dan Ma / Eric M Lynch / Scott E Boyken / Po-Ssu Huang / Lance Stewart / Frank DiMaio / Justin M Kollman / Ben F Luisi / Tomoaki Matsuura / William A Catterall / David Baker / |
PubMed 要旨 | Transmembrane channels and pores have key roles in fundamental biological processes and in biotechnological applications such as DNA nanopore sequencing, resulting in considerable interest in the ...Transmembrane channels and pores have key roles in fundamental biological processes and in biotechnological applications such as DNA nanopore sequencing, resulting in considerable interest in the design of pore-containing proteins. Synthetic amphiphilic peptides have been found to form ion channels, and there have been recent advances in de novo membrane protein design and in redesigning naturally occurring channel-containing proteins. 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. 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. |
リンク | Nature / PubMed:32848250 / PubMed Central |
手法 | EM (単粒子) / X線回折 |
解像度 | 2.4 - 7.6 Å |
構造データ | EMDB-20613, PDB-6u1s: EMDB-30126, PDB-6m6z: PDB-6o35: PDB-6tj1: PDB-6tms: |
化合物 | ChemComp-HOH: ChemComp-SO4: |
由来 |
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キーワード | DE NOVO PROTEIN (De novo) / nanopore / de novo design / MEMBRANE PROTEIN (膜タンパク質) / Helical bundle (ヘリックスバンドル) / octamer (オリゴマー) / computational design / pore / BIOSYNTHETIC PROTEIN (生合成) / hexamer (オリゴマー) / computational protein design / Transmembrane (膜貫通型タンパク質) / Rosetta |