scholarly journals An ECF-type transporter scavenges heme to overcome iron-limitation in Staphylococcus lugdunensis

eLife ◽  
2020 ◽  
Vol 9 ◽  
Author(s):  
Angelika Jochim ◽  
Lea Adolf ◽  
Darya Belikova ◽  
Nadine Anna Schilling ◽  
Inda Setyawati ◽  
...  
Keyword(s):  
Recombinant Expression ◽  
Iron Acquisition ◽  
Bacterial Pathogen ◽  
Energy Coupling ◽  
Coupling Factor ◽  
Iron Limitation ◽  
Staphylococcus Lugdunensis ◽  
Nutritional Limitation ◽  
Heme Acquisition ◽  
Factor Type

Energy-coupling factor type transporters (ECF) represent trace nutrient acquisition systems. Substrate binding components of ECF-transporters are membrane proteins with extraordinary affinity, allowing them to scavenge trace amounts of ligand. A number of molecules have been described as substrates of ECF-transporters, but an involvement in iron-acquisition is unknown. Host-induced iron limitation during infection represents an effective mechanism to limit bacterial proliferation. We identified the iron-regulated ECF-transporter Lha in the opportunistic bacterial pathogen Staphylococcus lugdunensis and show that the transporter is specific for heme. The recombinant substrate-specific subunit LhaS accepted heme from diverse host-derived hemoproteins. Using isogenic mutants and recombinant expression of Lha, we demonstrate that its function is independent of the canonical heme acquisition system Isd and allows proliferation on human cells as sources of nutrient iron. Our findings reveal a unique strategy of nutritional heme acquisition and provide the first example of an ECF-transporter involved in overcoming host-induced nutritional limitation.

scholarly journals Staphylococcus lugdunensis: a Skin Commensal with Invasive Pathogenic Potential

2020 ◽  
Vol 34 (2) ◽  
pp. e00205-20
Author(s):  
Simon Heilbronner ◽  
Timothy J. Foster
Keyword(s):  
Membrane Damage ◽  
Energy Coupling ◽  
Coupling Factor ◽  
Coagulase Negative Staphylococcus ◽  
Peptide Antibiotic ◽  
Staphylococcus Lugdunensis ◽  
Pathogenic Potential ◽  
Content Type ◽  
Flight Mass Spectrometry ◽  
Von Willebrand

SUMMARYStaphylococcus lugdunensis is a species of coagulase-negative staphylococcus (CoNS) that causes serious infections in humans akin to those of S. aureus. It was often misidentified as S. aureus, but this has been rectified by recent routine use of matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) in diagnostic laboratories. It encodes a diverse array of virulence factors for adhesion, cytotoxicity, and innate immune evasion, but these are less diverse than those encoded by S. aureus. It expresses an iron-regulated surface determinant (Isd) system combined with a novel energy-coupling factor (ECF) mechanism for extracting heme from hemoproteins. Small cytolytic S. lugdunensis synergistic hemolysins (SLUSH), peptides related to phenol-soluble modulins of S. aureus, act synergistically with β-toxin to lyse erythrocytes. S. lugdunensis expresses a novel peptide antibiotic, lugdunin, that can influence the nasal and skin microbiota. Endovascular infections are initiated by bacterial adherence to fibrinogen promoted by a homologue of Staphylococcus aureus clumping factor A and to von Willebrand factor on damaged endothelium by an uncharacterized mechanism. S. lugdunensis survives within mature phagolysosomes of macrophages without growing and is released only following apoptosis. This differs fundamentally from S. aureus, which actively grows and expresses bicomponent leukotoxins that cause membrane damage and could contribute to survival in the infected host. S. lugdunensis is being investigated as a probiotic to eradicate S. aureus from the nares of carriers. However, this is contraindicated by its innate virulence. Studies to obtain a deeper understanding of S. lugdunensis colonization, virulence, and microbiome interactions are therefore warranted.

Energy Coupling Factor-Type ABC Transporters for Vitamin Uptake in Prokaryotes

Biochemistry ◽  
2012 ◽  
Vol 51 (22) ◽  
pp. 4390-4396 ◽  
Author(s):  
Guus B. Erkens ◽  
Maria Majsnerowska ◽  
Josy ter Beek ◽  
Dirk Jan Slotboom
Keyword(s):  
Abc Transporters ◽  
Energy Coupling ◽  
Coupling Factor ◽  
Factor Type ◽  
Vitamin Uptake

scholarly journals An Iron-Regulated Autolysin Remodels the Cell Wall To Facilitate Heme Acquisition in Staphylococcus lugdunensis

2015 ◽  
Vol 83 (9) ◽  
pp. 3578-3589 ◽  
Author(s):  
Allison J. Farrand ◽  
Kathryn P. Haley ◽  
Nichole M. Lareau ◽  
Simon Heilbronner ◽  
John A. McLean ◽  
...  
Keyword(s):  
Cell Wall ◽  
Protein Localization ◽  
Iron Acquisition ◽  
Opportunistic Pathogen ◽  
System Component ◽  
Growth Defect ◽  
Staphylococcus Lugdunensis ◽  
Gene Encoding ◽  
Content Type ◽  
Heme Acquisition

Bacteria alter their cell surface in response to changing environments, including those encountered upon invasion of a host during infection. One alteration that occurs in several Gram-positive pathogens is the presentation of cell wall-anchored components of the iron-regulated surface determinant (Isd) system, which extracts heme from host hemoglobin to fulfill the bacterial requirement for iron.Staphylococcus lugdunensis, an opportunistic pathogen that causes infective endocarditis, encodes an Isd system. Unique among the known Isd systems,S. lugdunensiscontains a gene encoding a putative autolysin located adjacent to the Isd operon. To elucidate the function of this putative autolysin, here named IsdP, we investigated its contribution to Isd protein localization and hemoglobin-dependent iron acquisition.S. lugdunensisIsdP was found to be iron regulated and cotranscribed with the Isd operon. IsdP is a specialized peptidoglycan hydrolase that cleaves the stem peptide and pentaglycine crossbridge of the cell wall and alters processing and anchoring of a major Isd system component, IsdC. Perturbation of IsdC localization due toisdPinactivation results in a hemoglobin utilization growth defect. These studies establish IsdP as an autolysin that functions in heme acquisition and describe a role for IsdP in cell wall reorganization to accommodate nutrient uptake systems during infection.

scholarly journals Iron Limitation Triggers Early Egress by the Intracellular Bacterial Pathogen Legionella pneumophila

2016 ◽  
Vol 84 (8) ◽  
pp. 2185-2197 ◽  
Author(s):  
Tamara J. O'Connor ◽  
Huaixin Zheng ◽  
Susan M. VanRheenen ◽  
Soma Ghosh ◽  
Nicholas P. Cianciotto ◽  
...  
Keyword(s):  
Host Cell ◽  
Legionella Pneumophila ◽  
Iron Acquisition ◽  
Dominant Role ◽  
Bacterial Pathogen ◽  
Transport Protein ◽  
Iron Limitation ◽  
Growth Defect ◽  
Content Type

Legionella pneumophilais an intracellular bacterial pathogen that replicates in alveolar macrophages, causing a severe form of pneumonia. Intracellular growth of the bacterium depends on its ability to sequester iron from the host cell. In theL. pneumophilastrain 130b, one mechanism used to acquire this essential nutrient is the siderophore legiobactin. Iron-bound legiobactin is imported by the transport protein LbtU. Here, we describe the role of LbtP, a paralog of LbtU, in iron acquisition in theL. pneumophilastrain Philadelphia-1. Similar to LbtU, LbtP is a siderophore transport protein and is required for robust growth under iron-limiting conditions. Despite their similar functions, however, LbtU and LbtP do not contribute equally to iron acquisition. The Philadelphia-1 strain lacking LbtP is more sensitive to iron deprivationin vitro. Moreover, LbtP is important forL. pneumophilagrowth within macrophages while LbtU is dispensable. These results demonstrate that LbtP plays a dominant role over LbtU in iron acquisition. In contrast, loss of both LbtP and LbtU does not impairL. pneumophilagrowth in the amoebal hostAcanthamoeba castellanii, demonstrating a host-specific requirement for the activities of these two transporters in iron acquisition. The growth defect of the ΔlbtPmutant in macrophages is not due to alterations in growth kinetics. Instead, the absence of LbtP limitsL. pneumophilareplication and causes bacteria to prematurely exit the host cell. These results demonstrate the existence of a preprogrammed exit strategy in response to iron limitation that allowsL. pneumophilato abandon the host cell when nutrients are exhausted.

Discovery of Antibacterial Agents Inhibiting the Energy-Coupling Factor (ECF) Transporters by Structure-Based Virtual Screening

2020 ◽  
Author(s):  
Eleonora Diamanti ◽  
Inda Setyawati ◽  
Spyridon Bousis ◽  
leticia mojas ◽  
lotteke Swier ◽  
...  
Keyword(s):  
Virtual Screening ◽  
Antibacterial Agents ◽  
Antimicrobial Agents ◽  
Energy Coupling ◽  
Coupling Factor ◽  
Design Synthesis ◽  
Screening Design ◽  
Starting Point ◽  
Wide Range ◽  
Vitamin Uptake

Here, we report on the virtual screening, design, synthesis and structure–activity relationships (SARs) of the first class of selective, antibacterial agents against the energy-coupling factor (ECF) transporters. The ECF transporters are a family of transmembrane proteins involved in the uptake of vitamins in a wide range of bacteria. Inhibition of the activity of these proteins could reduce the viability of pathogens that depend on vitamin uptake. Because of their central role in the metabolism of bacteria and their absence in humans, ECF transporters are novel potential antimicrobial targets to tackle infection. The hit compound’s metabolic and plasma stability, the potency (20, MIC Streptococcus pneumoniae = 2 µg/mL), the absence of cytotoxicity and a lack of resistance development under the conditions tested here suggest that this scaffold may represent a promising starting point for the development of novel antimicrobial agents with an unprecedented mechanism of action.<br>

scholarly journals Competing for Iron: Duplication and Amplification of the isd Locus in Staphylococcus lugdunensis HKU09-01 Provides a Competitive Advantage to Overcome Nutritional Limitation

PLoS Genetics ◽  
2016 ◽  
Vol 12 (8) ◽  
pp. e1006246 ◽  
Author(s):  
Simon Heilbronner ◽  
Ian R. Monk ◽  
Jeremy R. Brozyna ◽  
David E. Heinrichs ◽  
Eric P. Skaar ◽  
...  
Keyword(s):  
Competitive Advantage ◽  
Staphylococcus Lugdunensis ◽  
Nutritional Limitation

scholarly journals Bacteriomimetic Liposomes Improve Antibiotic Activity of a Novel Energy-Coupling Factor Transporter Inhibitor

Pharmaceutics ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 4
Author(s):  
Menka Drost ◽  
Eleonora Diamanti ◽  
Kathrin Fuhrmann ◽  
Adriely Goes ◽  
Atanaz Shams ◽  
...  
Keyword(s):  
Model Organism ◽  
Energy Coupling ◽  
Coupling Factor ◽  
Free Drug ◽  
Drug Formulation ◽  
Phosphatidylcholine Liposomes ◽  
Loading Method ◽  
Fold Reduction ◽  
Hydrophobic Compound ◽  
Bacterial Growth Inhibition

Liposomes have been studied for decades as nanoparticulate drug delivery systems for cytostatics, and more recently, for antibiotics. Such nanoantibiotics show improved antibacterial efficacy compared to the free drug and can be effective despite bacterial recalcitrance. In this work, we present a loading method of bacteriomimetic liposomes for a novel, hydrophobic compound (HIPS5031) inhibiting energy-coupling factor transporters (ECF transporters), an underexplored antimicrobial target. The liposomes were composed of DOPG (18:1 (Δ9-cis) phosphatidylglycerol) and CL (cardiolipin), resembling the cell membrane of Gram-positive Staphylococcus aureus and Streptococcus pneumoniae, and enriched with cholesterol (Chol). The size and polydispersity of the DOPG/CL/± Chol liposomes remained stable over 8 weeks when stored at 4 °C. Loading of the ECF transporter inhibitor was achieved by thin film hydration and led to a high encapsulation efficiency of 33.19% ± 9.5% into the DOPG/CL/Chol liposomes compared to the phosphatidylcholine liposomes (DMPC/DPPC). Bacterial growth inhibition assays on the model organism Bacillus subtilis revealed liposomal HIPS5031 as superior to the free drug, showing a 3.5-fold reduction in CFU/mL at a concentration of 9.64 µM. Liposomal HIPS5031 was also shown to reduce B. subtilis biofilm. Our findings present an explorative basis for bacteriomimetic liposomes as a strategy against drug-resistant pathogens by surpassing the drug-formulation barriers of innovative, yet unfavorably hydrophobic, antibiotics.

scholarly journals A Novel Heme Transporter from the Energy Coupling Factor Family Is Vital for Group A Streptococcus Colonization and Infections

2020 ◽  
Vol 202 (14) ◽  
Author(s):  
Nilanjana Chatterjee ◽  
Laura C. C. Cook ◽  
Kristin V. Lyles ◽  
Hong Anh T. Nguyen ◽  
Darius J. Devlin ◽  
...  
Keyword(s):  
Inductively Coupled Plasma ◽  
Group A Streptococcus ◽  
Bacterial Colonization ◽  
Critical Role ◽  
Systemic Infection ◽  
Energy Coupling ◽  
Coupling Factor ◽  
Iron Complex ◽  
Invasive Infection ◽  
Group A

ABSTRACT Group A streptococcus (GAS) produces millions of infections worldwide, including mild mucosal infections, postinfection sequelae, and life-threatening invasive diseases. During infection, GAS readily acquires nutritional iron from host heme and hemoproteins. Here, we identified a new heme importer, named SiaFGH, and investigated its role in GAS pathophysiology. The SiaFGH proteins belong to a group of transporters with an unknown ligand from the recently described family of energy coupling factors (ECFs). A siaFGH deletion mutant exhibited high streptonigrin resistance compared to the parental strain, suggesting that iron ions or an iron complex is the likely ligand. Iron uptake and inductively coupled plasma mass spectrometry (ICP-MS) studies showed that the loss of siaFGH did not impact GAS import of ferric or ferrous iron, but the mutant was impaired in using hemoglobin iron for growth. Analysis of cells growing on hemoglobin iron revealed a substantial decrease in the cellular heme content in the mutant compared to the complemented strain. The induction of the siaFGH genes in trans resulted in the induction of heme uptake. The siaFGH mutant exhibited a significant impairment in murine models of mucosal colonization and systemic infection. Together, the data show that SiaFGH is a new type of heme importer that is key for GAS use of host hemoproteins and that this system is imperative for bacterial colonization and invasive infection. IMPORTANCE ECF systems are new transporters that take up various vitamins, cobalt, or nickel with a high affinity. Here, we establish the GAS SiaFGH proteins as a new ECF module that imports heme and demonstrate its importance in virulence. SiaFGH is the first heme ECF system described in bacteria. We identified homologous systems in the genomes of related pathogens from the Firmicutes phylum. Notably, GAS and other pathogens that use a SiaFGH-type importer rely on host hemoproteins for a source of iron during infection. Hence, recognizing the function of this noncanonical ABC transporter in heme acquisition and the critical role that it plays in disease has broad implications.

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