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 A Novel Class of Modular Transporters for Vitamins in Prokaryotes

2008 ◽  
Vol 191 (1) ◽  
pp. 42-51 ◽  
Author(s):  
Dmitry A. Rodionov ◽  
Peter Hebbeln ◽  
Aymerick Eudes ◽  
Josy ter Beek ◽  
Irina A. Rodionova ◽  
...  
Keyword(s):  
Abc Transporters ◽  
Transmembrane Protein ◽  
Energy Coupling ◽  
Coupling Factor ◽  
Human Pathogens ◽  
Gram Positive Bacteria ◽  
New Class ◽  
Vitamin Uptake ◽  
Genome Context ◽  
Nucleotide Binding Proteins

ABSTRACT The specific and tightly controlled transport of numerous nutrients and metabolites across cellular membranes is crucial to all forms of life. However, many of the transporter proteins involved have yet to be identified, including the vitamin transporters in various human pathogens, whose growth depends strictly on vitamin uptake. Comparative analysis of the ever-growing collection of microbial genomes coupled with experimental validation enables the discovery of such transporters. Here, we used this approach to discover an abundant class of vitamin transporters in prokaryotes with an unprecedented architecture. These transporters have energy-coupling modules comprised of a conserved transmembrane protein and two nucleotide binding proteins similar to those of ATP binding cassette (ABC) transporters, but unlike ABC transporters, they use small integral membrane proteins to capture specific substrates. We identified 21 families of these substrate capture proteins, each with a different specificity predicted by genome context analyses. Roughly half of the substrate capture proteins (335 cases) have a dedicated energizing module, but in 459 cases distributed among almost 100 gram-positive bacteria, including numerous human pathogens, different and unrelated substrate capture proteins share the same energy-coupling module. The shared use of energy-coupling modules was experimentally confirmed for folate, thiamine, and riboflavin transporters. We propose the name energy-coupling factor transporters for the new class of membrane transporters.

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 Two Essential Arginine Residues in the T Components of Energy-Coupling Factor Transporters

2009 ◽  
Vol 191 (21) ◽  
pp. 6482-6488 ◽  
Author(s):  
Olivia Neubauer ◽  
Anja Alfandega ◽  
Janna Schoknecht ◽  
Ulrich Sternberg ◽  
Anne Pohlmann ◽  
...  
Keyword(s):  
Atpase Activity ◽  
Transmembrane Protein ◽  
Energy Coupling ◽  
Coupling Factor ◽  
Single Mutation ◽  
Modular Assembly ◽  
Double Mutation ◽  
Arginine Residues ◽  
Uptake Activity ◽  
Vitamin Uptake

ABSTRACT Energy-coupling factor (ECF) transporters, a recently discovered class of importers of micronutrients, are composed of a substrate-specific transmembrane component (S component) and a conserved energy-coupling module consisting of a transmembrane protein (T component) and pairs of ABC ATPases (A proteins). Based on utilization of a dedicated (subclass I) or shared (subclass II) energy-coupling module, ECF systems fall into two subclasses. The T components are the least-characterized proteins of ECF importers, and their function is essentially unknown. Using RcBioN and LmEcfT, the T units of the subclass I biotin transporter (RcBioMNY) of a gram-negative bacterium and of the subclass II folate, pantothenate, and riboflavin transporters of a lactic acid bacterium, respectively, we analyzed the role of two strongly conserved short motifs, each containing an arginine residue. Individual replacement of the two Arg residues in RcBioN reduced ATPase activity, an indicator of the transporter function, by two-thirds without affecting the modular assembly of the RcBioMNY complex. A double Arg-to-Glu replacement destroyed the complex and abolished ATPase activity. The corresponding single mutation in motif II of LmEcfT, as well as a double mutation, led to loss of the T unit from the subclass II ECF transporters and inactivated these systems. A single Arg-to-Glu replacement in motif I, however, abolished vitamin uptake activity without affecting assembly of the modules. Our results indicate that the conserved motif I in T components is essential for intramolecular signaling and, in cooperation with motif II, for subunit assembly of modular ECF transporters.

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 Targeting the energy-coupling factor (ECF) transporters: identification of new tool compounds

2021 ◽  
Author(s):  
Eleonora Diamanti ◽  
Inda Setyawati ◽  
Spyridon Bousis ◽  
Paulo C. T. Souza ◽  
leticia mojas ◽  
...  
Keyword(s):  
Energy Coupling ◽  
Coupling Factor ◽  
Coarse Grained ◽  
Design Synthesis ◽  
Screening Design ◽  
Wide Range ◽  
Pharmaceutical Properties ◽  
Vitamin Uptake ◽  
Dynamics Simulations ◽  
Chemical Class

The energy-coupling factor (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. Their central role in the metabolism of bacteria and absence in humans make the ECF transporters a potential antibacterial target, which can be further investigated making use of a selective chemical probe. Here, we report on the virtual screening, design, synthesis, structure–activity relationships (SARs) and coarse-grained molecular dynamics simulations of the first class of inhibitors of the ECF transporters. We investigated the mechanism of action of this chemical class and profiled the best hit compounds regarding their pharmaceutical properties. The optimized hit has a minimum inhibitory concentration (MIC) value of 2 µg/mL against Streptococcus pneumoniae, which opens up the possibility to use this chemical class to investigate the role of the ECF transporters in health and disease.

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 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.

Structure and mechanism of energy-coupling factor transporters

2013 ◽  
Vol 21 (12) ◽  
pp. 652-659 ◽  
Author(s):  
Peng Zhang
Keyword(s):  
Energy Coupling ◽  
Coupling Factor
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