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TitleHelical reconstruction of and needle filaments attached to type 3 basal bodies.
Journal, issue, pagesBiochem Biophys Rep, Vol. 27, Page 101039, Year 2021
Publish dateJun 27, 2021
AuthorsVadim Kotov / Michele Lunelli / Jiri Wald / Michael Kolbe / Thomas C Marlovits /
PubMed AbstractGram-negative pathogens evolved a syringe-like nanomachine, termed type 3 secretion system, to deliver protein effectors into the cytoplasm of host cells. An essential component of this system is a ...Gram-negative pathogens evolved a syringe-like nanomachine, termed type 3 secretion system, to deliver protein effectors into the cytoplasm of host cells. An essential component of this system is a long helical needle filament that protrudes from the bacterial surface and connects the cytoplasms of the bacterium and the eukaryotic cell. Previous structural research was predominantly focused on reconstituted type 3 needle filaments, which lacked the biological context. In this work we introduce a facile procedure to obtain high-resolution cryo-EM structure of needle filaments attached to the basal body of type 3 secretion systems. We validate our approach by solving the structure of PrgI filament and demonstrate its utility by obtaining the first high-resolution cryo-EM reconstruction of Shigella MxiH filament. Our work paves the way to systematic structural characterization of attached type 3 needle filaments in the context of mutagenesis studies, protein structural evolution and drug development.
External linksBiochem Biophys Rep / PubMed:34258394 / PubMed Central
MethodsEM (helical sym.)
Resolution3.6 Å
Structure data

EMDB-11311, PDB-6znh:
Structure of the Salmonella PrgI needle filament attached to the basal body
Method: EM (helical sym.) / Resolution: 3.6 Å

EMDB-11312, PDB-6zni:
Structure of the Shigella MxiH needle filament attached to the basal body
Method: EM (helical sym.) / Resolution: 3.6 Å

Source
  • Salmonella enterica subsp. enterica serovar Typhimurium (bacteria)
  • salmonella typhimurium (bacteria)
  • shigella flexneri 5a str. m90t (bacteria)
KeywordsPROTEIN TRANSPORT / Type 3 secretion system / Salmonella / Helical reconstruction / Needle filament. / Shigella

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