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TitleTowards the application of Tc toxins as a universal protein translocation system.
Journal, issue, pagesNat Commun, Vol. 10, Issue 1, Page 5263, Year 2019
Publish dateNov 20, 2019
AuthorsDaniel Roderer / Evelyn Schubert / Oleg Sitsel / Stefan Raunser /
PubMed AbstractTc toxins are bacterial protein complexes that inject cytotoxic enzymes into target cells using a syringe-like mechanism. Tc toxins are composed of a membrane translocator and a cocoon that ...Tc toxins are bacterial protein complexes that inject cytotoxic enzymes into target cells using a syringe-like mechanism. Tc toxins are composed of a membrane translocator and a cocoon that encapsulates a toxic enzyme. The toxic enzyme varies between Tc toxins from different species and is not conserved. Here, we investigate whether the toxic enzyme can be replaced by other small proteins of different origin and properties, namely Cdc42, herpes simplex virus ICP47, Arabidopsis thaliana iLOV, Escherichia coli DHFR, Ras-binding domain of CRAF kinase, and TEV protease. Using a combination of electron microscopy, X-ray crystallography and in vitro translocation assays, we demonstrate that it is possible to turn Tc toxins into customizable molecular syringes for delivering proteins of interest across membranes. We also infer the guidelines that protein cargos must obey in terms of size, charge, and fold in order to apply Tc toxins as a universal protein translocation system.
External linksNat Commun / PubMed:31748551 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution2 - 5.1 Å
Structure data

EMDB-10314:
Tc holotoxin prepore TcdA1-TcdB2-TccC3-Cdc42
Method: EM (single particle) / Resolution: 5.1 Å

PDB-6sup:
Crystal Structure of TcdB2-TccC3-Cdc42
Method: X-RAY DIFFRACTION / Resolution: 2 Å

PDB-6suq:
Crystal Structure of TcdB2-TccC3-TEV
Method: X-RAY DIFFRACTION / Resolution: 3.7 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-HOH:
WATER

Source
  • photorhabdus luminescens (bacteria)
  • homo sapiens (human)
  • tobacco etch virus
KeywordsTOXIN / Toxins / Tc Toxins

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