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Defining the DmsA Signal Sequence Interaction with DmsD And TatBC

Ghosh, D.; Ramasamy, S. D.

2019-10-02 microbiology
10.1101/789909 bioRxiv
Show abstract

Redox - enzyme maturation proteins (REMP) ensure co-factor loading and folding of proteins targeted to the twin-arginine translocation (Tat) pathway. The details of the interaction of a REMP with the corresponding signal sequence of its substrate are not well understood. Here, we demonstrate the features of the signal sequence for the Tat substrate DmsA (ssDmsA) responsible for complex formation with its REMP, DmsD, and with the Tat membrane complex TatB & TatC (TatBC). A heterologously expressed ssDmsA/DmsD complex forms two stochiometric populations corresponding to monomeric and dimeric forms of the complex. The monomeric complex has a higher affinity for the TatBC complex than the dimeric, which imply higher level regulation process to ensure the maturation of protein before translocation. Results from various binding studies yielded the shortest signal peptide required for ssDmsA/DmsA interaction and the region responsible for the TatBC interaction. Further experiments like alanine scanning in this peptide highlight the possible residues that are essential for this complex formation.

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