Difference between revisions of "Part:BBa K554007"
Line 20: Line 20:
 
[[Image:UNICAMP_EMSE_secretion_device_schema.jpg|center|700px]]
 
[[Image:UNICAMP_EMSE_secretion_device_schema.jpg|center|700px]]
 
'''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.
 
'''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.
 +
 +
 +
=Three-dimensional structure representation==
 +
 +
You can find below a tridimensional structure of ATP-binding domain of hemolysin B from ''Escherichia coli'' (retrieved from PDB 1MT0 (Schmitt et al. 2003)) solved by X-ray crystallography at 2.5 A resolution. This is a jmol applet, in which you can interactively see the protein structure of the QseC cytoplasmic domain:
 +
<br>
 +
<div align="center">
 +
 +
<html>
 +
<script src="http://www.cbi.cnptia.embrapa.br/jmol/Jmol.js" type="text/javascript"></script>
 +
<script type="text/javascript">
 +
  jmolInitialize("http://www.cbi.cnptia.embrapa.br/jmol/");
 +
  jmolApplet(400, "load http://www.cbi.cnptia.embrapa.br/jmol/files/1mt0.pdb;");
 +
  jmolBr();
 +
 +
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")
 +
 +
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")
 +
 +
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")
 +
 +
  jmolButton("spin y" ,"spin y");
 +
  jmolButton("spin off" ,"spin off");
 +
</script>
 +
</div>
 +
<br />
 +
</html>
 +
 +
===References===
 +
 +
Schmitt, L.,  Benabdelhak, H.,  Blight, M.A.,  Holland, I.B.,  Stubbs, M.T.Crystal structure of the nucleotide-binding domain of the ABC-transporter haemolysin B: identification of a variable region within ABC helical domains.Journal: (2003) J.Mol.Biol. 330: 333-342 [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Abstract&list_uids=12823972 Link to PubMed]

Revision as of 05:06, 27 September 2011

Hemolysin B - HlyB

HlyB 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). HlyB acts as a ATP-binding cassette, and recognizes the substrate via its secretion signal (like HlyA) and is responsible for the specificity of the secretion system process. This system is composed of 4 essential parts: the C-terminal signal sequence of alpha-hemolysin (HlyA, which will be linked to the export target protein), the two specific translocator proteins HlyB and HlyD and the outer membrane protein [BBa_K554009 TolC].

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 1301
    Illegal BglII site found at 1964
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 231
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage and Biology

You can see a representation of this device acting in the schema below:

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.


Three-dimensional structure representation=

You can find below a tridimensional structure of ATP-binding domain of hemolysin B from Escherichia coli (retrieved from PDB 1MT0 (Schmitt et al. 2003)) solved by X-ray crystallography at 2.5 A resolution. This is a jmol applet, in which you can interactively see the protein structure of the QseC cytoplasmic domain:


References

Schmitt, L., Benabdelhak, H., Blight, M.A., Holland, I.B., Stubbs, M.T.Crystal structure of the nucleotide-binding domain of the ABC-transporter haemolysin B: identification of a variable region within ABC helical domains.Journal: (2003) J.Mol.Biol. 330: 333-342 [http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&dopt=Abstract&list_uids=12823972 Link to PubMed]