Difference between revisions of "Part:BBa K257007"

 
 
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__NOTOC__
 
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<partinfo>BBa_K257008 short</partinfo>
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<partinfo>BBa_K257007 short</partinfo>
  
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ClyA is exported to the periplasm and exposed in outer-membrane vesicles
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ClyA (or HlyE) is an alpha-PFT for Pore Forming Toxins.
  
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<!-- Add more about the biology of this part here
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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.
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*ClyA can be used to co-localize fully functional heterologous proteins directly in bacterial OMVs
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*We can fuse GFP to the C or N term of Cly A, to track OMVs easily.
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*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.
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*Cly A confers vesicle binding to and invasion of host cells.
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*ClyA was significantly enriched in OMVs relative to other lumenal and membrane bound OMV proteins.<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
  
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K257008 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K257007 SequenceAndFeatures</partinfo>
  
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  
 
===Functional Parameters===
 
===Functional Parameters===
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<partinfo>BBa_K257008 parameters</partinfo>
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<partinfo>BBa_K257007 parameters</partinfo>
 
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===References===
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<ol class="references">
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<li> The structure of a cytolytic a-helical toxin pore reveals its assembly mechanism, M.Mueller & N.Ban. 2009 - [http://www.ncbi.nlm.nih.gov/pubmed/19421192 | 19421192 ]</li>
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<li> 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. [http://www.ncbi.nlm.nih.gov/pubmed/14532000 14532000]</li>
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<li> 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. [http://www.ncbi.nlm.nih.gov/pubmed/11731136 11731136]</li>
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<li> 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.[http://www.ncbi.nlm.nih.gov/pubmed/15557633 15557633]</li>
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<li> 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. [http://www.ncbi.nlm.nih.gov/pubmed/18511069 18511069]</li>
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<li> 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.[http://www.ncbi.nlm.nih.gov/pubmed/14578354 14578354]</li>
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</ol>

Latest revision as of 07:06, 22 October 2009

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 - [http://www.ncbi.nlm.nih.gov/pubmed/19421192 | 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. [http://www.ncbi.nlm.nih.gov/pubmed/14532000 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. [http://www.ncbi.nlm.nih.gov/pubmed/11731136 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.[http://www.ncbi.nlm.nih.gov/pubmed/15557633 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. [http://www.ncbi.nlm.nih.gov/pubmed/18511069 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.[http://www.ncbi.nlm.nih.gov/pubmed/14578354 14578354]