Difference between revisions of "Part:BBa K554009"

 
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<partinfo>BBa_K554009 short</partinfo>
 
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TolC is ...
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TolC is part of the hemolysin secretion system ([http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Project#Device_3:_Secretion_system Device 3, Protein Secretion System]), very important to export the proteins produced inside bacteria and that must act in targets outside (such as [https://parts.igem.org/Part:BBa_K554005 IL-12] and [https://parts.igem.org/Part:BBa_K554004 IL-10]). TolC is a specific outer membrane protein, which forms a long channel throughout the outer membrane and the periplasm, largely open towards the extracellular medium.  
  
 
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<partinfo>BBa_K554009 parameters</partinfo>
 
<partinfo>BBa_K554009 parameters</partinfo>
 
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===Usage and Biology===
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'''You can see a representation of this device acting in the schema below:''' HlyB gene and product are shown as a symbolic cilinder in orange.
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[[Image:UNICAMP_EMSE_secretion_device_schema.jpg|center|700px]]
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'''Representation of device 3, the protein secretion system, in a Jedi bacteria that contains Device 1 (Adrenaline sensor/IL-12 producer). To export a protein, the bacteria must have the HlyD, HlyB and TolC proteins and the target protein must have a signal sequence (HlyA tail). In this case, the target protein to be secreted is IL-12.
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A more realistic schema of ABC transport system is shown below:
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[[Image:UNICAMP_EMSE_secretion_ABC.jpg|center|500px]]
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==Three-dimensional structure representation ==
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You can find below a tridimensional structure of Crystal structure of the open state of tolc outer membrane protein from ''Escherichia coli'' (retrieved from PDB 2VDD (Bavro et al. 2008)) solved by crystallography and X-ray diffraction at 3.3 A resolution. This is a jmol applet, in which you can interactively see the protein structure of TolC:
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<br>
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<div align="center">
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<html>
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<script src="http://www.cbi.cnptia.embrapa.br/jmol/Jmol.js" type="text/javascript"></script>
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<script type="text/javascript">
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  jmolInitialize("http://www.cbi.cnptia.embrapa.br/jmol/");
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  jmolApplet(400, "load http://www.cbi.cnptia.embrapa.br/jmol/files/2vdd.pdb;");
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  jmolBr();
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jmolButton("restrict not dna; restrict not protein; restrict not HOH; restrict not ligand; dots off; selectionhalos off; center selected; zoom 100; select all; wireframe; select all; color chain; select Cu or Cu1 or Zn or fe or Ca or Cl or Mn or Mg or na or Br or SO4 or ligand; color magenta selected; cpk; select all", "Stick view", "Stick view")
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jmolButton("restrict not HOH; select all; wireframe off;dots off; ribbon; color structure; zoom 100;  center selected; label off; backbone off; selectionhalos off; color structure; select Cu or Cu1 or Zn or fe or Ca or Cl or Mn or Mg or na or Br or SO4 or ligand; cpk; select ligand; wireframe 75; select all; Hbonds off; SSbonds off; select ligand; ribbon off", "Ribbon view", "Ribbon view")
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jmolButton("restrict not dna; restrict not protein; restrict not HOH; restrict not ligand; dots off; selectionhalos off; center selected; zoom 100; select all; wireframe; select all; color chain; backbone 50; wireframe off; select Cu or Cu1 or Zn or fe or Ca or Cl or Mn or Mg or na or Br or SO4 or ligand; cpk; select ligand; wireframe 75; select all; Hbonds off; SSbonds off; select ligand; backbone off", "Wireframe view", "Wireframe view")
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  jmolButton("spin y" ,"spin y");
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  jmolButton("spin off" ,"spin off");
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</script>
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</div>
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<br />
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</html>
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Molecule: OUTER MEMBRANE PROTEIN TOLC
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Polymer: 1 Type: protein Length: 460
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Chains: A, B, C
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Fragment: RESIDUES 1-450
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Mutation: YES
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UniProtKB: P02930
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===References===
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Barbara D. Tzschaschel, Carlos A. Guzmán,, Kenneth N. Timmis and Victor de Lorenzo. An Escherichia coli hemolysin transport system-based vector for the export of polypeptides: Export of shiga-like toxin IIeB subunit by Salmonella typhimurium aroA. Nature Biotechnology 14, 765 - 769 (1996) [http://www.nature.com/nbt/journal/v14/n6/abs/nbt0696-765.html Article link]
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P. Delepelaire. Type I secretion in Gram-negative bacteria.  Biochimia et Biophysica Actca 1694, 149-161 (2004) [http://www.ncbi.nlm.nih.gov/pubmed/15546664 Link to PubMed]
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Ivaylo Gentschev, Guido Dietrich and Werner Goebel. The E. coli α-hemolysin secretion system and its use in vaccine development. Trends in Microbiology 10, 39-45 (2002) [www.ncbi.nlm.nih.gov/pubmed/11755084 Link to PubMed]
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Bavro, V.N.,  Pietras, Z.,  Furnham, N.,  Perez-Cano, L.,  Fernandez-Recio, J.,  Pei, X.Y.,  Truer, R.,  Misra, R.,  Luisi, B.Assembly and channel opening in a bacterial drug efflux machine.Journal: (2008) Mol.Cell 30: 114 [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Abstract&list_uids=18406332 Link to PubMed]

Revision as of 12:45, 27 September 2011

TolC

TolC is part of the hemolysin secretion system ([http://2011.igem.org/Team:UNICAMP-EMSE_Brazil/Project#Device_3:_Secretion_system Device 3, Protein Secretion System]), very important to export the proteins produced inside bacteria and that must act in targets outside (such as IL-12 and IL-10). TolC is a specific outer membrane protein, which forms a long channel throughout the outer membrane and the periplasm, largely open towards the extracellular medium.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 395
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 247


Usage and Biology

You can see a representation of this device acting in the schema below: HlyB gene and product are shown as a symbolic cilinder in orange.

UNICAMP EMSE secretion device schema.jpg

Representation of device 3, the protein secretion system, in a Jedi bacteria that contains Device 1 (Adrenaline sensor/IL-12 producer). To export a protein, the bacteria must have the HlyD, HlyB and TolC proteins and the target protein must have a signal sequence (HlyA tail). In this case, the target protein to be secreted is IL-12.

A more realistic schema of ABC transport system is shown below:

UNICAMP EMSE secretion ABC.jpg


Three-dimensional structure representation

You can find below a tridimensional structure of Crystal structure of the open state of tolc outer membrane protein from Escherichia coli (retrieved from PDB 2VDD (Bavro et al. 2008)) solved by crystallography and X-ray diffraction at 3.3 A resolution. This is a jmol applet, in which you can interactively see the protein structure of TolC:


Molecule: OUTER MEMBRANE PROTEIN TOLC Polymer: 1 Type: protein Length: 460 Chains: A, B, C Fragment: RESIDUES 1-450 Mutation: YES UniProtKB: P02930


References

Barbara D. Tzschaschel, Carlos A. Guzmán,, Kenneth N. Timmis and Victor de Lorenzo. An Escherichia coli hemolysin transport system-based vector for the export of polypeptides: Export of shiga-like toxin IIeB subunit by Salmonella typhimurium aroA. Nature Biotechnology 14, 765 - 769 (1996) [http://www.nature.com/nbt/journal/v14/n6/abs/nbt0696-765.html Article link]

P. Delepelaire. Type I secretion in Gram-negative bacteria. Biochimia et Biophysica Actca 1694, 149-161 (2004) [http://www.ncbi.nlm.nih.gov/pubmed/15546664 Link to PubMed]

Ivaylo Gentschev, Guido Dietrich and Werner Goebel. The E. coli α-hemolysin secretion system and its use in vaccine development. Trends in Microbiology 10, 39-45 (2002) [www.ncbi.nlm.nih.gov/pubmed/11755084 Link to PubMed]

Bavro, V.N., Pietras, Z., Furnham, N., Perez-Cano, L., Fernandez-Recio, J., Pei, X.Y., Truer, R., Misra, R., Luisi, B.Assembly and channel opening in a bacterial drug efflux machine.Journal: (2008) Mol.Cell 30: 114 [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Abstract&list_uids=18406332 Link to PubMed]