Hughes, Adam M., Darby, John F. orcid.org/0000-0003-2754-6348, Dodson, Eleanor J. et al. (4 more authors) (2022) Peptide transport in Bacillus subtilis – structure and specificity in the extracellular solute binding proteins OppA and DppE. Microbiology (United Kingdom). 001274. ISSN 1465-2080
Abstract
Peptide transporters play important nutritional and cell signalling roles in Bacillus subtilis, which are pronounced during stationary phase adaptations and development. Three high-affinity ATP-binding cassette (ABC) family transporters are involved in peptide uptake – the oligopeptide permease (Opp), another peptide permease (App) and a less well-characterized dipeptide permease (Dpp). Here we report crystal structures of the extracellular substrate binding proteins, OppA and DppE, which serve the Opp and Dpp systems, respectively. The structure of OppA was determined in complex with endogenous peptides, mod-elled as Ser-Asn-Ser-Ser, and with the sporulation-promoting peptide Ser-Arg-Asn-Val-Thr, which bind with Kd values of 0.4 and 2 µM, respectively, as measured by isothermal titration calorimetry. Differential scanning fluorescence experiments with a wider panel of ligands showed that OppA has highest affinity for tetra-and penta-peptides. The structure of DppE revealed the unexpected presence of a murein tripeptide (MTP) ligand, l-Ala-d-Glu-meso-DAP, in the peptide binding groove. The mode of MTP binding in DppE is different to that observed in the murein peptide binding protein, MppA, from Escherichia coli, suggesting independent evolution of these proteins from an OppA-like precursor. The presence of MTP in DppE points to a role for Dpp in the uptake and recycling of cell wall peptides, a conclusion that is supported by analysis of the genomic context of dpp, which revealed adjacent genes encoding enzymes involved in muropeptide catabolism in a gene organization that is widely conserved in Firmicutes.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | Funding Information: This project was funded by the Biotechnology and Biological Sciences Research Council, UK, through a White Rose DTP Studentship to A.M.H. Acknowledgements We thank the Diamond Light Source for access to beamlines i02, i03 and i04 (proposal numbers mx-9948 and mx-13587) that contributed to the results presented here. Funding Information: This project was funded by the Biotechnology and Biological Sciences Research Council, UK, through a White Rose DTP Studentship to A.M.H. Publisher Copyright: © 2022 The Authors. |
Keywords: | Bacillus subtilis,DppE,murein peptide,OppA,peptide transport,sporulation |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Chemistry (York) The University of York > Faculty of Sciences (York) > Centre for Immunology and Infection (CII) (York) The University of York > Faculty of Sciences (York) > Biology (York) |
Funding Information: | Funder Grant number BBSRC (BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL) BB/T017805/1 |
Depositing User: | Pure (York) |
Date Deposited: | 12 Dec 2022 12:40 |
Last Modified: | 21 Jan 2025 18:06 |
Published Version: | https://doi.org/10.1099/mic.0.001274 |
Status: | Published |
Refereed: | Yes |
Identification Number: | 10.1099/mic.0.001274 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:194321 |