PDBsum entry 6tms

Biosynthetic protein

PDB id: 6tms

Contents

Protein chains

(+ 4 more) 69 a.a.

69 a.a.

Ligands

SER-GLY-LEU

HIS-HIS-HIS-HIS

SO4 ×5

Waters ×1077

PDB id:

6tms

Name:

Biosynthetic protein

Title:

Crystal structure of a de novo designed hexameric helical-bundle protein

Structure:

A novel designed pore protein. Chain: a, b, c, e, f, d, h, i, j, k. Engineered: yes. A novel designed pore protein. Chain: g, l. Engineered: yes. Affinity purification tag. Chain: q, r

Source:

Synthetic construct. Organism_taxid: 32630. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Organism_taxid: 32630

Resolution:

2.70Å R-factor: 0.296 R-free: 0.299

Authors:

C.Xu,X.Y.Pei,B.F.Luisi,D.Baker

Key ref:

C.Xu et al. (2020). Computational design of transmembrane pores. Nature, 585, 129-134. PubMed id: 32848250 DOI: 10.1038/s41586-020-2646-5

Date:

05-Dec-19 Release date: 29-Apr-20

Protein chains

No UniProt id for this chain

Struc: 69 a.a.

Protein chains

No UniProt id for this chain

Struc: 69 a.a.

DOI no: 10.1038/s41586-020-2646-5

Nature 585:129-134 (2020)

PubMed id: 32848250

Computational design of transmembrane pores.

C.Xu, P.Lu, T.M.Gamal El-Din, X.Y.Pei, M.C.Johnson, A.Uyeda, M.J.Bick, Q.Xu, D.Jiang, H.Bai, G.Reggiano, Y.Hsia, T.J.Brunette, J.Dou, D.Ma, E.M.Lynch, S.E.Boyken, P.S.Huang, L.Stewart, F.DiMaio, J.M.Kollman, B.F.Luisi, T.Matsuura, W.A.Catterall, D.Baker.

ABSTRACT

Transmembrane channels and pores have key roles in fundamental biological processes¹ 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.