Part:BBa_K4375012

Intimin domain for surface display (with anti-E epitope tag)

This protein contains the N-terminal domain of intimin, which is a bacterial adhesion factor, D0 an ig-like protein, and an E epitope tag. Intimin beta-barrel transmembrane domain anchors the construct into the outer membrane of the bacteria.

Usage and Biology

Intimin families are large, secreted polypeptides that contain three functional regions: i) a N-terminal signal peptide, that drives their Sec-dependent translocation across the inner membrane; ii) a β-domain, that is anchored into the outer membrane and comprises a 12-stranded β-barrel with a peptide linker running through the lumen of the β-barrel; and iii) a passenger region, that is secreted to the extracellular milieu. Although their mechanism of secretion remains uncertain, Intimin proteins are translocated into the periplasm and then use the β-barrel assembly machine (BAM) complex for insertion into the outer membrane and translocation of the passenger region to the cell surface. The passenger region is found in the C-terminus. The β-barrel of Intimin proteins is followed by a peptide linker that runs through the lumen connecting its C-terminus to the passenger region.

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 1983
  Illegal BamHI site found at 3999
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2004
  Illegal NgoMIV site found at 4020
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0075126#