PXD051962

PXD051962 is an original dataset announced via ProteomeXchange.

Title	E. coli acquires specific resistance to the antimicrobial peptide TAT-RasGAP (317-326) via modification of charge at the surface of the essential outer membrane insertase BamA
Description	Antimicrobial peptides (AMPs) are promising alternatives to classical antibiotics against multidrug 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 when passaged eight times with increasing concentrations of this peptide via mutations of the two-component system EnvZ/OmpR. To test whether additional mutations can cause further increase in resistance, we analysed a strain of E. coli showing high level of specific resistance to TAT-RasGAP (317-326) that we obtained after twenty passages of in vitro resistance selection. This strain bears, in addition to an envZ point mutation, a point mutation in bamA, an essential gene encoding an insertase involved in the insertion of outer membrane proteins. This mutation modifies the charge in a negatively charged loop (Q495-T505) at the surface of BamA. In silico docking simulations predict that binding affinity between TAT-RasGAP (317-326) and BamA varies depending on the charge of the Q495-T505 loop. We show here using CRISPR-Cas9-based targeted mutagenesis that mutations lowering the negative charge of the Q495-T505 loop decrease sensitivity of E. coli to TAT-RasGAP (317-326). Interestingly, BamA activity was not affected by TAT-RasGAP (317-326), indicating that BamA may function as a specific receptor for the AMP TAT-RasGAP (317-326). 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 multidrug resistant pathogens.
HostingRepository	PRIDE
AnnounceDate	2024-12-02
AnnouncementXML	Submission_2024-12-02_07:22:35.832.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 Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2024-05-03 05:29:06	ID requested
⏵ 1	2024-12-02 07:22:36	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. PXD051962
     1. Label: PRIDE project
     2. Name: E. coli acquires specific resistance to the antimicrobial peptide TAT-RasGAP (317-326) via modification of charge at the surface of the essential outer membrane insertase BamA