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TitleStructure of the calcium-dependent type 2 secretion pseudopilus.
Journal, issue, pagesNat Microbiol, Vol. 2, Issue 12, Page 1686-1695, Year 2017
Publish dateOct 9, 2017
AuthorsAracelys López-Castilla / Jenny-Lee Thomassin / Benjamin Bardiaux / Weili Zheng / Mangayarkarasi Nivaskumar / Xiong Yu / Michael Nilges / Edward H Egelman / Nadia Izadi-Pruneyre / Olivera Francetic /
PubMed AbstractMany Gram-negative bacteria use type 2 secretion systems (T2SSs) to secrete proteins involved in virulence and adaptation. Transport of folded proteins via T2SS nanomachines requires the assembly of ...Many Gram-negative bacteria use type 2 secretion systems (T2SSs) to secrete proteins involved in virulence and adaptation. Transport of folded proteins via T2SS nanomachines requires the assembly of inner membrane-anchored fibres called pseudopili. Although efficient pseudopilus assembly is essential for protein secretion, structure-based functional analyses are required to unravel the mechanistic link between these processes. Here, we report an atomic model for a T2SS pseudopilus from Klebsiella oxytoca, obtained by fitting the NMR structure of its calcium-bound subunit PulG into the ~5-Å-resolution cryo-electron microscopy reconstruction of assembled fibres. This structure reveals the comprehensive network of inter-subunit contacts and unexpected features, including a disordered central region of the PulG helical stem, and highly flexible C-terminal residues on the fibre surface. NMR, mutagenesis and functional analyses highlight the key role of calcium in PulG folding and stability. Fibre disassembly in the absence of calcium provides a basis for pseudopilus length control, essential for protein secretion, and supports the Archimedes screw model for the type 2 secretion mechanism.
External linksNat Microbiol / PubMed:28993624 / PubMed Central
MethodsEM (helical sym.) / NMR (solution)
Resolution5.0 Å
Structure data

8812

5wda

EMDB-8812, PDB-5wda:
Structure of the PulG pseudopilus
Method: EM (helical sym.) / Resolution: 5.0 Å

PDB-5o2y:
NMR structure of the calcium bound form of PulG, major pseudopilin from Klebsiella oxytoca T2SS
Method: SOLUTION NMR

Chemicals

ChemComp-CA:
Unknown entry

Source
  • klebsiella oxytoca (bacteria)
KeywordsPROTEIN TRANSPORT / Klebsiella oxytoca T2SS / major pseudopilin / calcium / helical polymer / bacterial secretion / cryo-EM

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