Journal: Nat Struct Mol Biol / Year: 2016 Title: Membrane insertion of a Tc toxin in near-atomic detail. Authors: Christos Gatsogiannis / Felipe Merino / Daniel Prumbaum / Daniel Roderer / Franziska Leidreiter / Dominic Meusch / Stefan Raunser / Abstract: Tc toxins from pathogenic bacteria use a special syringe-like mechanism to perforate the host cell membrane and inject a deadly enzyme into the host cytosol. The molecular mechanism of this unusual ...Tc toxins from pathogenic bacteria use a special syringe-like mechanism to perforate the host cell membrane and inject a deadly enzyme into the host cytosol. The molecular mechanism of this unusual injection system is poorly understood. Using electron cryomicroscopy, we determined the structure of TcdA1 from Photorhabdus luminescens embedded in lipid nanodiscs. In our structure, compared with the previous structure of TcdA1 in the prepore state, the transmembrane helices rearrange in the membrane and open the initially closed pore. However, the helices do not span the complete membrane; instead, the loops connecting the helices form the rim of the funnel. Lipid head groups reach into the space between the loops and consequently stabilize the pore conformation. The linker domain is folded and packed into a pocket formed by the other domains of the toxin, thereby considerably contributing to stabilization of the pore state.
Num. of particles selected: 30061 / Details: particles were picked manually
Symmetry
Point symmetry: C5 (5 fold cyclic)
3D reconstruction
Resolution: 3.46 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 13000 Details: The density was filtered to its local resolution, using localfilt (SPARX) and RESMAP Symmetry type: POINT
Atomic model building
Protocol: AB INITIO MODEL
Refinement
Highest resolution: 3.46 Å
Refine LS restraints
Refine-ID
Type
Dev ideal
Number
ELECTRONMICROSCOPY
f_bond_d
0.013
11395
ELECTRONMICROSCOPY
f_angle_d
1.127
15475
ELECTRONMICROSCOPY
f_dihedral_angle_d
11.213
9277
ELECTRONMICROSCOPY
f_chiral_restr
0.059
1708
ELECTRONMICROSCOPY
f_plane_restr
0.007
2018
+
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