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bioRxiv · 10.1101/261537

Mutation in DsbA signal sequence hampers the SRP mechanism: A new strategy to combat virulence factor

Durrani, F. G.·Gul, R.·Mirza, M. U.·Kaderbhai, N. N.·Saleem, M.·Akhter, M. W.
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Abstract

Disulphide bond (Dsb) protein, characterized as an important virulence factor in gram negative bacteria. In this study, amino acid mutations in DsbA signal sequence (ss) and its effect on translocation of recombinant Ovine growth hormone (rOGH) was observed. Eight constructs were designed on the basis of increased hydrophobicity and showed that hydrophobicity and specificity of amino acid plays a crucial role in translocation of rOGH. Two DsbAss with the same hydropathy (1.539), one had alteration at -13 and second at -11 position; alanine (Ala) to isoleucine respectively were designed. The former DsbAss translocated rOGH from membrane to cytoplasmic fraction in E. coli as confirmed by SDS-PAGE, Western blot and molecular modelling analysis. MD simulations and binding free energy calculations evidenced that, altering Ala changed the orientation of signal peptide in the Ffh-M domain binding groove and hampered the process of translocation while change at position -11 pointed it outward. We hypothesize, amino acid and position of mutations in DsbAss can hinder the translocation process of signal recognition particle system, thus affecting the virulence of bacteria.

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Durrani, F. G., Gul, R., Mirza, M. U., Kaderbhai, N. N., Saleem, M., Akhter, M. W.. 2018-02-07. Mutation in DsbA signal sequence hampers the SRP mechanism: A new strategy to combat virulence factor. https://doi.org/10.1101/261537

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