Details
| Original language | English |
|---|---|
| Pages (from-to) | 1134-1146 |
| Number of pages | 13 |
| Journal | ACS infectious diseases |
| Volume | 8 |
| Issue number | 6 |
| Early online date | 2 May 2022 |
| Publication status | Published - 10 Jun 2022 |
Abstract
The development of new antibiotics against Gram-negative bacteria has to deal with the low permeability of the outer membrane. This obstacle can be overcome by utilizing siderophore-dependent iron uptake pathways as entrance routes for antibiotic uptake. Iron-chelating siderophores are actively imported by bacteria, and their conjugation to antibiotics allows smuggling the latter into bacterial cells. Synthetic siderophore mimetics based on MECAM (1,3,5-N,N′,N″-tris-(2,3-dihydroxybenzoyl)-triaminomethylbenzene) and DOTAM (1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane) cores, both chelating iron via catechol groups, have been recently applied as versatile carriers of functional cargo. In the present study, we show that MECAM and the MECAM-ampicillin conjugate 3 transport iron into Pseudomonas aeruginosa cells via the catechol-type outer membrane transporters PfeA and PirA and DOTAM solely via PirA. Differential proteomics and quantitative real-time polymerase chain reaction (qRT-PCR) showed that MECAM import induced the expression of pfeA, whereas 3 led to an increase in the expression of pfeA and ampc, a gene conferring ampicillin resistance. The presence of DOTAM did not induce the expression of pirA but upregulated the expression of two zinc transporters (cntO and PA0781), pointing out that bacteria become zinc starved in the presence of this compound. Iron uptake experiments with radioactive 55Fe demonstrated that import of this nutrient by MECAM and DOTAM was as efficient as with the natural siderophore enterobactin. The study provides a functional validation for DOTAM- and MECAM-based artificial siderophore mimetics as vehicles for the delivery of cargo into Gram-negative bacteria.
Keywords
- antibiotic vectorization, iron uptake, outer membrane transporters, Pseudomonas aeruginosa, siderophores, Trojan horse
ASJC Scopus subject areas
- Medicine(all)
- Infectious Diseases
Sustainable Development Goals
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In: ACS infectious diseases, Vol. 8, No. 6, 10.06.2022, p. 1134-1146.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Uptake Mechanisms and Regulatory Responses to MECAM- and DOTAM-Based Artificial Siderophores and Their Antibiotic Conjugates in Pseudomonas aeruginosa
AU - Fritsch, Sarah
AU - Gasser, Véronique
AU - Peukert, Carsten
AU - Pinkert, Lukas
AU - Kuhn, Lauriane
AU - Perraud, Quentin
AU - Normant, Vincent
AU - Brönstrup, Mark
AU - Schalk, Isabelle J.
N1 - Funding Information: This work was partially funded by the Centre National de la Recherche Scientifique, a grant from the Joint Programming Initiative on Antimicrobial Resistance (JPI AMR, grant number: 01KI1825) and the Interdisciplinary Thematic Institute (ITI) InnoVec (Innovative Vectorization of Biomolecules, IdEx, ANR-10-IDEX-0002) and SFRI (ANR-20-SFRI-0012). The mass spectrometry instrumentation at the IBMC was funded by the University of Strasbourg, IdEx “Equipement mi-lourd” 2015. The equipment at the IPHC was partly funded by the French Proteomics Infrastructure (ProFI; ANR-10-INSB-08-03). Q.P. had a fellowship from the associations Vaincre la Mucoviscidose and Gregory Lemarchal. C.P. had a Kekulé stipend from the ‘Verband der chemischen Industrie,’ and L.P. was funded by the DFG project number BR 3572/4-1.
PY - 2022/6/10
Y1 - 2022/6/10
N2 - The development of new antibiotics against Gram-negative bacteria has to deal with the low permeability of the outer membrane. This obstacle can be overcome by utilizing siderophore-dependent iron uptake pathways as entrance routes for antibiotic uptake. Iron-chelating siderophores are actively imported by bacteria, and their conjugation to antibiotics allows smuggling the latter into bacterial cells. Synthetic siderophore mimetics based on MECAM (1,3,5-N,N′,N″-tris-(2,3-dihydroxybenzoyl)-triaminomethylbenzene) and DOTAM (1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane) cores, both chelating iron via catechol groups, have been recently applied as versatile carriers of functional cargo. In the present study, we show that MECAM and the MECAM-ampicillin conjugate 3 transport iron into Pseudomonas aeruginosa cells via the catechol-type outer membrane transporters PfeA and PirA and DOTAM solely via PirA. Differential proteomics and quantitative real-time polymerase chain reaction (qRT-PCR) showed that MECAM import induced the expression of pfeA, whereas 3 led to an increase in the expression of pfeA and ampc, a gene conferring ampicillin resistance. The presence of DOTAM did not induce the expression of pirA but upregulated the expression of two zinc transporters (cntO and PA0781), pointing out that bacteria become zinc starved in the presence of this compound. Iron uptake experiments with radioactive 55Fe demonstrated that import of this nutrient by MECAM and DOTAM was as efficient as with the natural siderophore enterobactin. The study provides a functional validation for DOTAM- and MECAM-based artificial siderophore mimetics as vehicles for the delivery of cargo into Gram-negative bacteria.
AB - The development of new antibiotics against Gram-negative bacteria has to deal with the low permeability of the outer membrane. This obstacle can be overcome by utilizing siderophore-dependent iron uptake pathways as entrance routes for antibiotic uptake. Iron-chelating siderophores are actively imported by bacteria, and their conjugation to antibiotics allows smuggling the latter into bacterial cells. Synthetic siderophore mimetics based on MECAM (1,3,5-N,N′,N″-tris-(2,3-dihydroxybenzoyl)-triaminomethylbenzene) and DOTAM (1,4,7,10-tetrakis(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane) cores, both chelating iron via catechol groups, have been recently applied as versatile carriers of functional cargo. In the present study, we show that MECAM and the MECAM-ampicillin conjugate 3 transport iron into Pseudomonas aeruginosa cells via the catechol-type outer membrane transporters PfeA and PirA and DOTAM solely via PirA. Differential proteomics and quantitative real-time polymerase chain reaction (qRT-PCR) showed that MECAM import induced the expression of pfeA, whereas 3 led to an increase in the expression of pfeA and ampc, a gene conferring ampicillin resistance. The presence of DOTAM did not induce the expression of pirA but upregulated the expression of two zinc transporters (cntO and PA0781), pointing out that bacteria become zinc starved in the presence of this compound. Iron uptake experiments with radioactive 55Fe demonstrated that import of this nutrient by MECAM and DOTAM was as efficient as with the natural siderophore enterobactin. The study provides a functional validation for DOTAM- and MECAM-based artificial siderophore mimetics as vehicles for the delivery of cargo into Gram-negative bacteria.
KW - antibiotic vectorization
KW - iron uptake
KW - outer membrane transporters
KW - Pseudomonas aeruginosa
KW - siderophores
KW - Trojan horse
UR - http://www.scopus.com/inward/record.url?scp=85130030556&partnerID=8YFLogxK
U2 - 10.1021/acsinfecdis.2c00049
DO - 10.1021/acsinfecdis.2c00049
M3 - Article
C2 - 35500104
AN - SCOPUS:85130030556
VL - 8
SP - 1134
EP - 1146
JO - ACS infectious diseases
JF - ACS infectious diseases
SN - 2373-8227
IS - 6
ER -