The Bacillus subtilis TatAdCd system exhibits an extreme level of substrate selectivity

Frain, Kelly M and Jones, Alexander S. and Schoner, Ronald and Walker, Kelly L. and Robinson, Colin (2016) The Bacillus subtilis TatAdCd system exhibits an extreme level of substrate selectivity. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1864 (1). pp. 202-208. ISSN 0167-4889. (doi:https://doi.org/10.1016/j.bbamcr.2016.10.018) (Access to this publication is currently restricted. You may be able to access a copy if URLs are provided)

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Abstract

The Tat system preferentially transports correctly folded proteins across the bacterial membrane although little is known of the proofreading mechanism. Most research has focused on TatABC systems from Gram-negative bacteria, especially Escherichia coli, and much less is known of the TatAC-type systems from Gram-positive organisms. We have previously shown that the Bacillus subtilis TatAdCd system is functional in an E. coli tat null background and able to transport TorA-GFP and native TorA (TMAO reductase); here, we examined its ability to transport other proteins bearing a TorA signal sequence. We show that whereas E. coli TatABC transports a wide range of biotherapeutics including human growth hormone, interferon α2b, a VH domain protein and 2 different scFvs, TatAdCd transports the scFvs but completely rejects the other proteins. The system also rejects two native E. coli substrates, NrfC and FhuD. Moreover, we have shown that TatABC will transport a wide range of folded scFv variants with the surface altered to incorporate multiple salt bridges, charged residues (5 glutamate, lysine or arginine), or hydrophobic residues (up to 6 leucines). In contrast, TatAdCd completely rejects many of these variants including those with 5 or 6 added Leu residues. The combined data show that the TatABC and TatAdCd systems have very different substrate selectivities, with the TatAdCd system displaying an extreme level of selectivity when compared to the E. coli system. The data also provide a preliminary suggestion that TatAdCd may not tolerate surface domains with a level of hydrophobicity above a certain threshold.

Item Type: Article
Subjects: Q Science > QH Natural history
Q Science > QH Natural history > QH301 Biology
Divisions: Faculties > Sciences > School of Biosciences
Depositing User: Colin Robinson
Date Deposited: 12 Dec 2016 09:55 UTC
Last Modified: 23 Jan 2017 15:13 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/59622 (The current URI for this page, for reference purposes)
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