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Computational design of transmembrane pores

DOI: 10.1038/s41586-020-2646-5 DOI Help

Authors: Chunfu Xu (University of Washington) , Peilong Lu (University of Washington; Westlake University; Westlake Institute for Advanced Study) , Tamer M. Gamal El-Din (University of Washington) , Xue Y. Pei (University of Cambridge) , Matthew C. Johnson (University of Washington) , Atsuko Uyeda (Osaka University) , Matthew J. Bick (University of Washington) , Qi Xu (Westlake University; Westlake Institute for Advanced Study) , Daohua Jiang (University of Washington) , Hua Bai (University of Washington) , Gabriella Reggiano (University of Washington) , Yang Hsia (University of Washington) , T. J. Brunette (University of Washington) , Jiayi Dou (University of Washington) , Dan Ma (Westlake University; Westlake Institute for Advanced Study) , Eric M. Lynch (University of Washington) , Scott E. Boyken (University of Washington) , Po-Ssu Huang (Stanford University; University of Washington) , Lance Stewart (University of Washington) , Frank Dimaio (University of Washington) , Justin M. Kollman (University of Washington) , Ben Luisi (University of Cambridge) , Tomoaki Matsuura (Osaka University) , William A. Catterall (University of Washington) , David Baker (University of Washington)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature , VOL 585 , PAGES 129 - 134

State: Published (Approved)
Published: September 2020
Diamond Proposal Number(s): 9537

Abstract: 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.

Journal Keywords: Membrane proteins; Membrane structure and assembly; Protein design; Protein folding

Subject Areas: Biology and Bio-materials, Chemistry

Instruments: I02-Macromolecular Crystallography

Other Facilities: Proxima-2A beamline at SOLEIL; Advanced Light Source

Added On: 28/09/2020 14:45

Discipline Tags:

Biochemistry Chemistry Structural biology Biophysics Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)