Part:BBa_K4015002

DsbA

DsbA is a soluble periplasmic protein. It belongs to the thioredoxin family and acts as a type of signal peptide which serve as the primary source of disulfide bonds to secreted proteins. The protein catalyzes the oxidation of the two cysteine sulfhydryls and form a covalent S-S bond connecting amino acid, then donates to the secreted protein, catalyzing their oxidation and their folding. DsbA is primarily active in type II secretion system.

Usage and Biology

In our project, we pick three signal peptides to help secrete our target proteins, which are HlyA、GeneIII and DsbA . We used p47 vector containing GFP to compare the effectiveness of these signal peptides. p47-GFP-HlyA displayed a lower level of fluorescence comparing to p47-GeneIII-GFP, suggesting HlyA is more effective in secretion since more GFP has entered the medium. p47-DsbA-GFP showed no fluorescence, meaning protein expression was inhibited or the folding of the protein has failed. 

 Fig. Result of GFP secretion experiment 

Citation:

1.  Linton, E., Walsh, M. K., Sims, R. C., & Miller, C. D. (2011). Translocation of green fluorescent protein by comparative analysis with multiple signal peptides. Biotechnology Journal, 7(5), 667-676. https://doi.org/10.1002/biot.201100158 

2. Santos-Martin, C., Wang, G., Subedi, P., Hor, L., Totsika, M., Paxman, J. J., & Heras, B. (2021). Structural bioinformatic analysis of DsbA proteins and their pathogenicity associated substrates. Computational and Structural Biotechnology Journal, 19, 4725-4737. https://doi.org/10.1016/j.csbj.2021.08.018

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]