 |
PDBsum entry 6tms
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Biosynthetic protein
|
PDB id
|
|
|
|
6tms
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
Contents |
 |
|
|
|
|
|
|
|
|
|
|
|
(+ 4 more)
69 a.a.
|
|
|
|
|
|
|
|
|
|
|
69 a.a.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
References listed in PDB file
|
|
|
Key reference
|
|
|
Title
|
|
Computational design of transmembrane pores.
|
|
|
Authors
|
|
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.
|
|
|
Ref.
|
|
Nature, 2020,
585,
129-134.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
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.
|
|
|
|
|
|
|
