The outer membrane usher forms a twin-pore secretion complex.
The PapC usher is an outer membrane protein required for assembly and secretion of P pili in uropathogenic Escherichia coli. P pilus biogenesis occurs by the chaperone/usher pathway, a terminal branch of the general secretory pathway. Periplasmic chaperone-subunit complexes target to the PapC usher for fiber assembly and secretion through the usher to the cell surface. The molecular details of pilus biogenesis at the usher, and protein secretion across the outer membrane in general, are unclear. We studied the structure and oligomeric state of PapC by gel filtration, dynamic light scattering, and electron microscopy and image analysis. Two-dimensional crystals of wild-type PapC and a C-terminal deletion mutant of PapC were produced by reconstituting detergent purified usher into E.coli lipids. PapC formed a dimer both in detergent solution and in the phospholipid bilayer. Cryo-electron microscopy revealed that the usher forms a twin-pore complex. Removal of the C-terminal domain did not change the basic shape of the PapC molecule, but altered the dimeric association of the usher, suggesting that the C terminus forms part of the dimerization interface. The overall molecular size (11 nm), pore size (2 nm), and twin-pore configuration of PapC resemble that of the Tom40 complex, a mitochondrial outer membrane protein translocase.
Li
Huilin
H
Biology Department, Brookhaven National Laboratory, 50 Bell Ave, Upton, NY 11973, USA.
Qian
Luping
L
Chen
Zhiqiang
Z
Thibault
Danielle
D
Liu
Guang
G
Liu
Tianbo
T
Thanassi
David G
DG
D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism*; D003460:Crystallization; D003902:Detergents / Q000494:pharmacology; D019281:Dimerization; D004926:Escherichia coli / Q000737:chemistry / Q000166:cytology / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008854:Microscopy, Electron; D009154:Mutation / Q000235:genetics; D018272:Porins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D017384:Sequence Deletion / Q000235:genetics; D012996:Solutions / Q000737:chemistry; D013816:Thermodynamics
D003902:Detergents; D029968:Escherichia coli Proteins; D008051:Lipid Bilayers; D018272:Porins; D012996:Solutions; C111243:atpG protein, E coli