Part:BBa_K3016100

Vibrio natriegens' TorA signal peptide

This part contains Vibrio natriegens' TorA Tat signal peptide. It is a twin-arginine (RR) motif containing signal peptide for periplasmic transport of proteins via the twin-arginine translocation (Tat) pathway. Derived from Vibrio natriegens’ torA gene.

Biology

The twin-arginine translocation (Tat) pathway is capable of translocating fully folded proteins up to 150 kDa. It also contains a quality control feature of rejecting misfolded proteins. In some cases, disulfide bridge formation is not required for successful translocation. (Alanen et al., 2015)

Translocation using the tat-pathway requires the protein to contain a N-terminal signal peptide with a twin-arginine (RR) motif. The signal peptide is cleaved during the translocation process. A pair of V. natriegens’ native twin-arginine signal peptides identified by Aalto-Helsinki can be found here (TorAand Aminotransferase)

Use

This part can be N-terminally fused with your protein of choice to facilitate its periplasmic transport via the twin-arginine translocation (Tat) pathway.

Characterization

Aalto-Helsinki 2019 characterized this part by combining it with YGFP (BBa_K3016600), a slow-bleaching GFP variant, creating a composite TorA-YGFP part (BBa_K3016200) to test protein translocation into Vibrio natriegens' and Escherichia coli's periplasm.

File:TorA-YGFP natriegens.png

"Vibrio natriegens cells expressing TorA-YGFP

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]

References:

Alanen, H. I., Walker, K. L., Suberbie, M. L. V., Matos, C. F., Bönisch, S., Freedman, R. B., ... & Robinson, C. (2015). Efficient export of human growth hormone, interferon α2b and antibody fragments to the periplasm by the Escherichia coli Tat pathway in the absence of prior disulfide bond formation. Biochimica et Biophysica Acta (BBA)-Molecular Cell Research, 1853(3), 756-763.