Designing a novel signal peptide for secretion of recombinant Human activin A protein through the twin-arginine translocation (Tat)pathway in E. coli

One of the most prominent features of the E. coli, which makes it widely used in the production of recombinant proteins, is the ability of secretion of the desired protein into the extracellular space. The twin-arginine pathway (Tat), due to the possibility of secretion of the recombinant protein in the folded state, has a high potential for facilitating downstream processes of restoring the 3Dstructure and hence the function of proteins. In this project, regarding the key role of signal sequences in bacterial protein secretion, we gathered the signal sequences from two Archaea Haloferax volcanii and Halobacterium salinarumand E. coli itself from the UniProt database and then the eight sequences were aligned by Clustal Omega software. The final sequence with conserved and common amino acids was obtained. To confirm the secretion of human activin A linked to the designed signal peptide, further studies were carried out with TatP, PRED-Tat, and Phobius software, all of which are signal peptide indicative tools. The output scores, in addition to confirming the ability of the designed signal peptide in Human Active A secretion, indicating its specificity for the Tat pathway. The key step in translocation of protein to the extracellular space is the identification of the signal sequence by its receptor on the inner surface of the membrane. Comparison of the interaction energy of the designed signal peptide and its receptor with the homologous in E.coli with the foldX software showed more efficient bacterial secretion.

کلیدواژه ها:

The twin-arginine translocation (Tat) ، Human Activin A ، Signal peptide