Coding
clyA_Nter

Part:BBa_K257007

Designed by: Guillaume Cambray   Group: iGEM09_Paris   (2009-09-10)

ClyA_Fusion_N-Term

ClyA (or HlyE) is an alpha-PFT for Pore Forming Toxins.

PFTs are potent virulence factors class starting in a soluble form to an outer membrane-integrated pore. They exhibit their toxic effect either by membrane permeability barrier destruction or by toxic components delivery through the pores which forming by several assembly 8 or 13 ClyA subunits.

  • ClyA can be used to co-localize fully functional heterologous proteins directly in bacterial OMVs
  • We can fuse GFP to the C or N term of Cly A, to track OMVs easily.
  • ClyA is capable of co-localizing a variety of structurally diverse fusion partners to the surface of E. coli and their released vesicles, but only when the periplasmic disulfide bond-forming machinery was present ,it’s makes OMVs an ideal structure to transport hydrophobic compounds like membrane proteins into the host.
  • Cly A confers vesicle binding to and invasion of host cells.
  • ClyA was significantly enriched in OMVs relative to other lumenal and membrane bound OMV proteins.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 84
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

  1. The structure of a cytolytic a-helical toxin pore reveals its assembly mechanism, M.Mueller & N.Ban. 2009 - | 19421192
  2. S.N. Wai, B.Lindmark, T.Soderblom, A.Takade, M.Westermark, J.Oscarsson, et al. Vesicle-mediated export and assembly of pore-forming oligomers of the enterobacterial ClyA cytotoxin, 2003, Cell, 115, 25–35. 14532000
  3. F.J del Castillo, F. Moreno. and I.del Castillo. Secretion of the Escherichia coli K-12 SheA hemolysin is independent of its cytolytic activity, 2001, FEMS Microbiol.Lett. 204, 281–285. 11731136
  4. J.E.Galen, L.Zhao, M.Chinchilla, J.Y.Wang,M.F.Pasetti, J.Green and M.M. Levine. Adaptation of the endogenous Salmonella enterica serovar Typhi clyA-encoded hemolysin for antigen export enhances the immunogenicity of anthrax protective antigen domain 4 expressed by the attenuated live-vector vaccine strain CVD 908-htrA, 2004, Infect. Immun. 72, 7096–7106.15557633
  5. J.Y. Kim, A.M. Doody, D. J. Chen, G.H. Cremona, M.L. Shuler, D.Putnam,and M.P. DeLisa.Engineered. Bacterial Outer Membrane Vesicles with Enhanced Functionality, 2008, J. Mol. Biol. 380, 51–66. 18511069
  6. N.C.Kesty, M.J.Kuehn. Incorporation of heterologous outer membrane and periplasmic proteins into Escherichia coli outer membrane vesicles, 2004, J Biol Chem. 279(3):2069-76.14578354
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