Coding

Part:BBa_K554007

Designed by: UNICAMP EMSE Brazil team   Group: iGEM11_UNICAMP-EMSE_Brazil   (2011-09-21)
Revision as of 12:44, 27 September 2011 by Neshich.iap (Talk | contribs) (→‎Three-dimensional structure representation)

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


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]

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]
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