PXD055766

PXD055766 is an original dataset announced via ProteomeXchange.

Title	High level resistance to the antimicrobial peptide TAT-RasGAP (317-326) results from sequential acquisition of mutations influencing the Escherichia coli cell envelope biogenesis and composition
Description	Antimicrobial peptides (AMPs) are promising alternatives to classical antibiotics against antibiotic-resistant pathogens. TAT-RasGAP (317-326) is an AMP with broad range antibacterial activity, but its mechanism of action is unknown. Escherichia coli can become partially resistant to TAT-RasGAP (317-326) in vitro. In this study, we analysed a strain of E. coli with extensive resistance to TAT-RasGAP (317-326) but not to other AMPs that we obtained after twenty passages during an in vitro resistance selection experiment. This strain accumulates four mutations, that all apparently affect bacterial envelope composition. One of these mutations is a point mutation in bamA, which encodes an essential protein involved in the folding and proper insertion of outer membrane proteins. The point mutation resulted in a charge change in an area of the protein corresponding to a negatively charged loop at the surface of BamA. Using CRISPR-Cas9-based targeted mutagenesis, we showed that mutations lowering the negative charge of this loop decreased sensitivity of E. coli to TAT-RasGAP (317-326). In silico simulations unveiled the molecular driving forces responsible for the interaction between TAT-RasGAP (317-326) and BamA. These results indicated that electrostatic interactions, particularly hydrogen bonds, are involved in the stability of the molecular complex, representing a predictive fingerprint of the TAT-RasGAP (317-326) - BamA interaction strength. Interestingly, BamA activity was not affected by TAT-RasGAP (317-326), indicating that BamA may function as a specific receptor for this AMP. Our results indicate that binding and entry of TAT-RasGAP (317-326) may involve different mechanisms compared to other AMPs, which is in line with limited cross-resistance observed between different AMPs. This limited cross-resistance is important for the clinical application of AMPs towards drug-resistant pathogens.
HostingRepository	PRIDE
AnnounceDate	2024-12-02
AnnouncementXML	Submission_2024-12-02_07:16:01.992.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Nicolas Jacquier
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2024-09-10 04:07:39	ID requested
⏵ 1	2024-12-02 07:16:02	announced

10.1016/J.JBC.2024.108018;

submitter keyword: resistance,Antimicrobial peptides, outer membrane, Gram-negative bacteria

Nicolas Jacquier
contact affiliation	University Hospital Center and University of Lausanne, Institute of Microbiology, Bugnon 48, 1011 Lausanne
contact email	nicolas.jacquier@chuv.ch
lab head
Nicolas Jacquier
contact affiliation	Institute of Microbiology, University Hospital Center and University of Lausanne
contact email	nicolas.jacquier@chuv.ch
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD055766
     1. Label: PRIDE project
     2. Name: High level resistance to the antimicrobial peptide TAT-RasGAP (317-326) results from sequential acquisition of mutations influencing the Escherichia coli cell envelope biogenesis and composition