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TitleVirB8 homolog TraE from plasmid pKM101 forms a hexameric ring structure and interacts with the VirB6 homolog TraD.
Journal, issue, pagesProc Natl Acad Sci U S A, Vol. 115, Issue 23, Page 5950-5955, Year 2018
Publish dateJun 5, 2018
AuthorsBastien Casu / Charline Mary / Aleksandr Sverzhinsky / Aurélien Fouillen / Antonio Nanci / Christian Baron /
PubMed AbstractType IV secretion systems (T4SSs) are multiprotein assemblies that translocate macromolecules across the cell envelope of bacteria. X-ray crystallographic and electron microscopy (EM) analyses have ...Type IV secretion systems (T4SSs) are multiprotein assemblies that translocate macromolecules across the cell envelope of bacteria. X-ray crystallographic and electron microscopy (EM) analyses have increasingly provided structural information on individual T4SS components and on the entire complex. As of now, relatively little information has been available on the exact localization of the inner membrane-bound T4SS components, notably the mostly periplasmic VirB8 protein and the very hydrophobic VirB6 protein. We show here that the membrane-bound, full-length version of the VirB8 homolog TraE from the plasmid pKM101 secretion system forms a high-molecular-mass complex that is distinct from the previously characterized periplasmic portion of the protein that forms dimers. Full-length TraE was extracted from the membranes with detergents, and analysis by size-exclusion chromatography, cross-linking, and size exclusion chromatography (SEC) multiangle light scattering (MALS) shows that it forms a high-molecular-mass complex. EM and small-angle X-ray scattering (SAXS) analysis demonstrate that full-length TraE forms a hexameric complex with a central pore. We also overproduced and purified the VirB6 homolog TraD and show by cross-linking, SEC, and EM that it binds to TraE. Our results suggest that TraE and TraD interact at the substrate translocation pore of the secretion system.
External linksProc Natl Acad Sci U S A / PubMed:29784815 / PubMed Central
MethodsSAS (X-ray synchrotron)
Structure data

SASDB75:
Escherichia coli TraE protein: A VirB8 homolog from plasmid pKM101
Method: SAXS/SANS

Source
  • Escherichia coli (E. coli)

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