ATP synthase subunit c (atpE) is a transmembrane component of the F<sub>O</sub> sector of ATP synthase, critical for proton translocation and ATP production in Bacillus cereus. Recombinant versions, such as those expressed in E. coli with N-terminal His tags, enable studies on bacterial energetics and drug discovery .
Recombinant atpE is produced in E. coli via plasmid expression, followed by affinity chromatography using nickel-NTA resins due to the His tag . Key steps include:
Lyophilization: Stabilized with trehalose for long-term storage .
Reconstitution: Requires deionized water and glycerol (5–50%) to maintain solubility .
Forms the rotor (c-ring) of F<sub>O</sub> sector, coupling proton gradient to ATP synthesis .
Mutations in atpE (e.g., A17S in S. aureus) disrupt ATP production and confer antibiotic resistance .
B. cereus Rex protein modulates atpE expression under anaerobic conditions, linking oxygen availability to metabolic flux .
Tomatidine (TO) and derivatives inhibit ATP synthase by binding atpE, with resistance linked to atpE mutations (e.g., A17S) .
Bacillus ATP synthase is structurally conserved, making atpE a candidate for cross-species drug development .
Deletion of rex in B. cereus alters carbon flux through lactate pathways, indirectly affecting ATP synthase activity .
KEGG: bcg:BCG9842_B5519
ATP synthase subunit c (atpE) in Bacillus cereus is a critical component of the F₀ sector of ATP synthase, the enzyme complex responsible for ATP production through oxidative phosphorylation. It forms an oligomeric ring in the membrane that acts as a rotor, facilitating proton translocation across the membrane, which drives the conformational changes necessary for ATP synthesis. In Bacillales like B. cereus, ATP synthase also plays an important role in pH homeostasis and is essential for normal growth and cellular metabolism. The protein contains 72 amino acids forming a highly hydrophobic structure that spans the membrane, with a sequence of MSLGVIAAAIAIGLSALGAGIGNGLIVSRTIEGVARQPELKGALQTIMFIGVALVEALPII GVVIAFIVMNK .
The Bacillus cereus ATP synthase subunit c (atpE) consists of 72 amino acids organized into a predominantly hydrophobic structure. This protein forms two transmembrane α-helices connected by a polar loop region . Multiple copies of subunit c arrange in a ring formation within the membrane, creating a rotating element that channels protons across the membrane. The conserved carboxylate residue (typically a glutamate or aspartate) in one of the transmembrane helices is essential for proton translocation, functioning as the site of protonation/deprotonation during rotation. The highly conserved nature of these structural elements across Bacillales reflects the fundamental importance of this protein in energy metabolism.
ATP synthase subunit c (atpE) functions as part of the membrane-embedded F₀ sector of the ATP synthase complex. During ATP synthesis, protons flow through a channel at the interface between subunit a and the c-ring, causing the c-ring to rotate. Each subunit c contains a critical carboxylate residue that alternately binds and releases protons as it rotates through different environments. This rotation is mechanically coupled to the central stalk of the F₁ sector, inducing conformational changes in the catalytic sites that drive ATP synthesis. The number of c subunits in the ring determines the H⁺/ATP ratio, directly affecting the bioenergetic efficiency of the organism. In Bacillales, proper functioning of this complex is particularly crucial, as studies have shown that ATP synthase plays multiple roles beyond energy production, including pH homeostasis .
For effective expression of recombinant Bacillus cereus ATP synthase subunit c (atpE), several systems can be employed with specific optimizations:
E. coli Expression Systems:
Specialized strains: C41(DE3) or C43(DE3), designed specifically for membrane protein expression
Vector selection: pET series vectors with T7 promoter systems for controlled expression
Fusion tags: MBP, SUMO, or Thioredoxin tags to enhance solubility of this hydrophobic protein
Expression conditions: Induction at lower temperatures (16-20°C) with reduced IPTG concentrations (0.1-0.5 mM)
Cell-Free Expression Systems:
Particularly valuable for membrane proteins like atpE
Allows direct incorporation into nanodiscs or liposomes
Eliminates toxicity issues that may occur in cellular systems
Bacillus Expression Systems:
Homologous expression in Bacillus subtilis provides native-like membrane environment
May facilitate proper folding and assembly compared to heterologous systems
The choice of expression system should be guided by the intended experimental applications, with careful optimization of induction conditions, temperature, and extraction methods to maintain the native conformation of this highly hydrophobic membrane protein.
A multi-step purification strategy is recommended for obtaining high-yield, high-purity recombinant Bacillus cereus ATP synthase subunit c (atpE) while preserving its native conformation:
Membrane Protein Extraction:
Use gentle detergents like n-dodecyl-β-D-maltoside (DDM) or digitonin
Buffer composition: 20 mM Tris-HCl (pH 8.0), 150 mM NaCl, and 1-2% detergent
Include protease inhibitors to prevent degradation
Affinity Chromatography:
Employ a suitable affinity tag (His-tag is commonly used)
Wash extensively to remove nonspecifically bound proteins
Elute with imidazole gradient or specific protease cleavage
Size Exclusion Chromatography:
Separate oligomeric forms and remove aggregates
Evaluate protein quality by monitoring elution profile
Ion Exchange Chromatography:
Further enhance purity based on charge differences
Select appropriate resin based on predicted isoelectric point
Storage Considerations:
This comprehensive purification approach ensures that the protein maintains its structural integrity while achieving research-grade purity levels.
The hydrophobic nature of ATP synthase subunit c (atpE) presents significant challenges during purification. Researchers can employ the following strategies to overcome these challenges:
Optimized Detergent Selection:
Systematic screening of detergents (DDM, LMNG, digitonin)
Detergent concentration optimization to prevent aggregation while avoiding excess micelles
Consideration of novel amphipathic polymers like SMALPs that extract membrane proteins with their native lipid environment
Lipid Supplementation:
Addition of specific phospholipids during purification to maintain native-like environment
Use of lipid nanodiscs as a stabilizing platform for structural and functional studies
Stabilizing Additives:
Temperature Management:
Conducting all purification steps at 4°C to reduce dynamics and aggregation
Controlled cooling rates during storage preparation
Chimeric Constructs and Fusion Partners:
Design of chimeric constructs with soluble proteins
Selection of fusion partners that enhance expression and solubility while allowing native folding
Alternative Purification Approaches:
Co-purification with interacting partners or antibodies
Use of styrene-maleic acid copolymers to extract membrane proteins with their native lipid environment
These approaches, often used in combination, can significantly improve the yield and quality of purified atpE protein for downstream structural and functional studies.
Several functional assays can be employed to evaluate the activity of recombinant Bacillus cereus ATP synthase subunit c (atpE):
ATP Synthesis Assay:
Using inverted membrane vesicles containing reconstituted ATP synthase complexes
ATP production measured by luciferase-based luminescence detection
Allows correlation between ATP synthase activity and antibiotic potency
Can be performed with varying substrate concentrations to determine kinetic parameters
Proton Translocation Assay:
Employs pH-sensitive fluorescent dyes (ACMA, pyranine, or SNARF-1)
Monitors proton movement across membranes containing reconstituted ATP synthase
Provides direct evidence of atpE's role in proton transport
ATP Hydrolysis Assay:
Measures the reverse reaction (ATP hydrolysis)
Utilizes coupled enzyme systems (pyruvate kinase/lactate dehydrogenase)
Tracks NADH oxidation spectrophotometrically at 340 nm
Provides insights into the bidirectional functionality of the complex
Inhibitor Binding Studies:
Isothermal titration calorimetry (ITC) for thermodynamic parameters
Surface plasmon resonance (SPR) for binding kinetics
Fluorescence-based assays for high-throughput screening of potential inhibitors
Reconstitution into Proteoliposomes:
Incorporation of purified atpE into artificial liposomes
Assessment of proton pumping capabilities
Evaluation of the minimal system required for function
These assays collectively provide comprehensive insights into the functional integrity and activity of recombinant Bacillus cereus atpE in different experimental contexts.
To effectively study interactions between Bacillus cereus ATP synthase subunit c (atpE) and potential antimicrobial compounds, researchers should employ a multi-faceted approach:
Binding Assays:
Isothermal Titration Calorimetry (ITC): Provides direct measurement of binding thermodynamics and stoichiometry
Surface Plasmon Resonance (SPR): Offers real-time binding kinetics
Microscale Thermophoresis (MST): Works with small sample amounts and native conditions
Functional Inhibition Assays:
ATP Synthesis Inhibition: Using membrane vesicle assays to measure IC50 values for compounds
Growth Inhibition: Determining MIC values against wild-type and atpE mutant strains
Comparative analysis between bacterial ATP synthase inhibition and mitochondrial ATP synthase inhibition to assess selectivity index (as seen with FC04-100, which showed >10^5-fold selectivity)
Structural Analysis:
Molecular docking simulations
Hydrogen-deuterium exchange mass spectrometry to identify binding interfaces
Structural studies using NMR or X-ray crystallography if possible
Resistance Development Analysis:
Site-directed mutagenesis to create atpE variants for testing specific binding site hypotheses
This comprehensive approach provides both biophysical evidence of direct interaction and functional evidence of the biological relevance of such interactions.
Visualizing atpE oligomerization and c-ring formation requires specialized techniques that can capture these molecular assemblies. The following approaches provide complementary information about this critical aspect of ATP synthase structure:
Cryo-Electron Microscopy (cryo-EM):
Single-particle analysis can achieve near-atomic resolution of membrane protein complexes
Captures c-rings in their native oligomeric state without crystallization
Can visualize the complete ATP synthase complex including the c-ring
Blue Native PAGE:
Preserves native protein complexes during electrophoresis
Can separate different oligomeric states
When combined with western blotting, specifically identifies atpE-containing complexes
Chemical Cross-linking Coupled with Mass Spectrometry:
Covalently links adjacent subunits in the c-ring
Identifies interaction interfaces between subunits
Provides stoichiometric information about the complex
Fluorescence Resonance Energy Transfer (FRET):
Labels atpE with donor and acceptor fluorophores
Measures energy transfer as an indication of proximity
Can monitor assembly dynamics in real-time
Atomic Force Microscopy (AFM):
Visualizes membrane-embedded c-rings at nanometer resolution
Provides topographical information about ring height and diameter
Can be performed in near-native conditions
Native Mass Spectrometry:
Determines the intact mass of the c-ring complex
Provides precise stoichiometric information
Can detect bound lipids and small molecules
Electron Crystallography:
Applicable to 2D crystals of membrane proteins
Has been successfully used for c-rings from other bacterial species
Provides detailed structural information
These complementary approaches together provide comprehensive insights into the assembly, structure, and dynamics of atpE oligomerization and c-ring formation in Bacillus cereus ATP synthase.
Critical amino acid residues in Bacillus cereus ATP synthase subunit c (atpE) and their functional importance include:
Conserved Carboxylate Residue:
Polar/Charged Residues in Transmembrane Regions:
Rare in transmembrane domains but crucial for proton pathways
Create hydrophilic environment for proton movement
Mutations disrupt proton translocation without affecting assembly
Glycine-Rich Motifs:
Patterns like GXXXGXXXG facilitate helix-helix packing in the c-ring
Enable tight packing of transmembrane helices
Mutations destabilize the oligomeric structure
Critical for maintaining proper c-ring architecture
Drug-Binding Site Residues:
Interface Residues with Other Subunits:
N and C-terminal regions interact with other ATP synthase components
Alterations disrupt the assembly of the complete complex
Essential for energy coupling between F₀ and F₁ sectors
Experimental studies with ATP synthase inhibitors have demonstrated that mutations conferring resistance often come at a functional cost, reducing ATP production capacity as observed in S. aureus SCV strains , highlighting the critical nature of these residues.
Mutations in the atpE gene profoundly affect the sensitivity of Bacillus cereus to ATP synthase inhibitors through several mechanisms:
Altered Binding Site Architecture:
Functional Consequences:
Differential Effects on Inhibitor Classes:
Structural Impact on c-ring:
Mutations may alter c-ring assembly or stability
Changes in proton translocation pathways can affect inhibitor access
Alterations in c-ring rotation mechanics can impact inhibitor binding dynamics
Species-Specific Effects:
Understanding these structure-function relationships is crucial for designing next-generation ATP synthase inhibitors that can overcome or bypass resistance mechanisms in B. cereus and related pathogens.
The relationship between atpE mutations, ATP synthesis capacity, and bacterial fitness represents a complex interplay with significant implications for bacterial physiology and antimicrobial resistance:
Functional Trade-offs:
Mutations in atpE that confer resistance to inhibitors frequently reduce ATP synthase efficiency
Studies with tomatidine-resistant S. aureus mutants demonstrated that atpE mutations further reduced ATP production compared to parent strains
This suggests a direct molecular cost of resistance at the enzyme level
Compensatory Metabolic Adaptations:
Growth Rate and Competitive Fitness:
Reduced ATP synthesis typically correlates with decreased growth rates
In competitive environments, resistant mutants may be outcompeted by wild-type strains
This fitness cost can influence the persistence of resistant strains in the absence of selection pressure
Physiological Consequences Beyond Energy Production:
Environmental Context Dependence:
The fitness cost of atpE mutations may vary according to environmental conditions
In energy-limited environments, the cost of reduced ATP synthesis is likely magnified
Under certain stress conditions, reduced metabolism might paradoxically increase survival
Evolutionary Implications:
The fitness cost creates selective pressure for compensatory mutations
Secondary mutations may restore fitness while maintaining resistance
This evolutionary pathway influences the stability of resistance in bacterial populations
This complex relationship highlights the importance of considering both the direct antimicrobial effects and the evolutionary consequences when targeting ATP synthase as an antimicrobial strategy.
Mutations in the atpE gene contribute to antimicrobial resistance through multiple mechanisms:
Understanding these resistance mechanisms at the molecular level is crucial for developing effective antimicrobial strategies targeting ATP synthase in Bacillus cereus and related pathogens.
Several strategic approaches can be employed to overcome resistance to ATP synthase inhibitors targeting Bacillus cereus atpE:
Structural Modification of Inhibitors:
Develop derivatives with enhanced binding properties, as demonstrated with FC04-100, a tomatidine derivative that prevents high-level resistance development
Design compounds that interact with multiple sites on atpE simultaneously
Optimize inhibitor molecules to maintain high selectivity indices (bacterial vs. mitochondrial ATP synthase inhibition)
Combination Therapy Approaches:
Use ATP synthase inhibitors alongside antibiotics with different targets
Implement dual-targeting molecules that interact with atpE and another essential bacterial protein
Develop inhibitors that target multiple subunits of the ATP synthase complex simultaneously
Resistance-Resistant Design:
Target highly conserved, functionally essential residues where mutations would severely impair function
Design inhibitors that bind to regions requiring multiple simultaneous mutations for resistance
Target interfaces between subunits rather than individual subunits
Alternative Delivery Mechanisms:
Develop prodrug approaches that evade efflux mechanisms
Utilize nanoparticle-based delivery to increase local concentration
Implement membrane-penetrating peptides to enhance delivery
Predictive Resistance Modeling:
Employ computational approaches to predict potential resistance mutations
Preemptively design inhibitors effective against predicted resistant variants
Utilize directed evolution studies to identify potential resistance pathways
Metabolic Sensitization:
Target alternative energy production pathways alongside ATP synthase
Create conditions where ATP synthase function becomes even more critical
Identify synthetic lethal interactions with atpE mutations
This multi-faceted approach to inhibitor design and deployment can significantly reduce the probability of resistance development while maintaining therapeutic efficacy against Bacillus cereus infections.
The high conservation of ATP synthase subunit c (atpE) across Bacillales has profound implications for antimicrobial development strategies:
Narrow-Spectrum Targeting Opportunity:
The conserved sequence within Bacillales enables development of order-specific antibiotics
Tomatidine and its analog FC04-100 demonstrate this principle with their narrow yet specific spectrum of activity against SCVs of Bacillales
This allows targeting pathogenic Bacillales while potentially sparing beneficial microbiota
Conservation of Critical Functional Elements:
Highly conserved residues often indicate functional importance and limited mutational tolerance
Targeting these conserved elements increases the barrier to resistance development
The shared sequence identity suggests that findings from one species may translate to others within the order
Predictable Cross-Species Efficacy:
Inhibitors effective against atpE in one Bacillales species likely work across the order
This enables more efficient drug development with broader application within the target group
Conservation patterns can guide rational design of broad-spectrum (within Bacillales) inhibitors
Resistance Mechanism Anticipation:
Conservation allows prediction of resistance mechanisms across species
Mutations conferring resistance in one species may indicate potential resistance paths in related organisms
This knowledge facilitates proactive inhibitor design to counter anticipated resistance
Evolutionary Constraints:
Structural Targeting Precision:
Understanding and leveraging this conservation pattern is crucial for developing effective and targeted antimicrobials against Bacillus cereus and other clinically relevant members of the Bacillales order.
Several cutting-edge structural biology techniques show exceptional promise for elucidating the high-resolution structure and dynamics of Bacillus cereus ATP synthase subunit c (atpE):
Cryo-Electron Microscopy (Cryo-EM):
Single-particle analysis can achieve near-atomic resolution of membrane proteins without crystallization
Developments in direct electron detectors and data processing algorithms have dramatically improved resolution
Can capture different conformational states relevant to the functional cycle
Particularly suitable for visualizing the entire ATP synthase complex including the c-ring
Cryo-Electron Tomography (Cryo-ET) with Subtomogram Averaging:
Allows visualization of ATP synthase complexes directly in cellular membranes
Preserves native lipid interactions and supramolecular arrangements
Combined with focused ion beam milling to visualize proteins in their cellular context
Integrative Structural Biology Approaches:
Combining multiple experimental techniques (X-ray, NMR, SAXS) with computational modeling
Cross-validation between methods enhances confidence in structural details
Particularly powerful for membrane protein complexes like ATP synthase
Native Mass Spectrometry:
Determines intact mass and stoichiometry of membrane protein complexes
Reveals lipid-protein interactions preserved during ionization
Provides insights into c-ring assembly and stability
Solid-State NMR Spectroscopy:
Can resolve atomic-level details of membrane proteins in lipid bilayers
Particularly informative for dynamics and protonation states
Dynamic Nuclear Polarization (DNP) enhances sensitivity for difficult samples
Serial Femtosecond Crystallography with X-ray Free Electron Lasers (XFEL):
"Diffraction before destruction" approach overcomes radiation damage limitations
Can use microcrystals grown in lipidic environments
Potential to capture transient structural states during the catalytic cycle
These advanced techniques, especially when used in combination, offer unprecedented potential to reveal the structure of B. cereus atpE in its functional context, providing critical insights for rational drug design targeting this essential component.
Systems biology approaches offer powerful frameworks for understanding the broader metabolic consequences of ATP synthase subunit c (atpE) function and inhibition in Bacillus cereus:
Multi-omics Integration:
Combining transcriptomics, proteomics, and metabolomics data from B. cereus under varying energy conditions
Integrating temporal dynamics of metabolic shifts following ATP synthase modulation
Constructing comprehensive models of energy metabolism regulation
Metabolic Flux Analysis:
¹³C-based flux analysis to track metabolic pathways dependent on ATP synthase activity
Quantification of flux redistribution through central carbon metabolism under ATP limitation
Identification of alternate ATP generation pathways activated when ATP synthase function is compromised
Genome-Scale Metabolic Modeling:
Construction of genome-scale metabolic models incorporating ATP synthase constraints
In silico prediction of growth phenotypes under varying ATP synthase activity levels
Identification of synthetic lethal targets that could be inhibited alongside atpE
Network Analysis:
Protein-protein interaction network analysis to identify functional modules connected to ATP synthase
Regulatory network mapping to understand transcriptional responses to energy limitation
Identification of network hubs and vulnerabilities in ATP-limited conditions
Comparative Systems Analysis:
Cross-species analysis comparing ATP synthase-dependent metabolic networks
Assessment of differential impacts between pathogenic B. cereus and beneficial microbes
Identification of species-specific vulnerabilities in energy metabolism
Experimental Validation Approaches:
Construction of reporter strains to monitor real-time changes in energy status
Targeted metabolite analysis focusing on energy-related compounds (ATP/ADP ratio, NADH/NAD⁺)
Creation of conditional knockdowns to mimic inhibitor effects with temporal control
Research has already provided evidence for metabolic remodeling in persistent bacteria and SCVs , suggesting that systems approaches would reveal valuable insights into how B. cereus responds to energy limitation induced by atpE inhibition or mutation.
Advanced computational approaches offer powerful tools for predicting both inhibitor binding to Bacillus cereus atpE and potential resistance development:
Molecular Docking and Virtual Screening:
Predicts binding modes and affinities of potential inhibitors to atpE
Enables virtual screening of large compound libraries
Incorporates flexible docking to account for protein adaptability
Allows structure-based design of optimized inhibitors
Molecular Dynamics (MD) Simulations:
Models the dynamic behavior of atpE-inhibitor complexes in a membrane environment
Identifies stable binding conformations and key interaction residues
Reveals potential water-mediated interactions and conformational changes upon binding
Typically runs for hundreds of nanoseconds to microseconds to capture relevant dynamics
Quantum Mechanics/Molecular Mechanics (QM/MM):
Provides detailed insights into electronic properties of binding interactions
Particularly valuable for understanding protonation states and hydrogen bonding networks
Can model reaction mechanisms relevant to inhibitor binding
Machine Learning Approaches:
Develops predictive models for binding affinity based on structural and chemical features
Identifies patterns in existing inhibitor data to guide new compound design
Can predict cross-resistance patterns between structurally related compounds
Resistance Prediction Methods:
In silico mutagenesis to systematically assess the impact of potential resistance mutations
Prediction of mutational hotspots based on sequence conservation and structural constraints
Estimation of fitness costs associated with resistance mutations
Molecular dynamics simulations of mutant proteins to assess structural and functional consequences
Evolutionary Algorithms and Network Models:
Simulates evolutionary pathways to resistance under different selection pressures
Models the probability of specific resistance mutations emerging
Predicts compensatory mutations that might restore fitness in resistant strains
Identifies optimal inhibitor properties to minimize resistance development
Integration of these computational approaches with experimental validation creates a powerful platform for rational design of ATP synthase inhibitors with reduced potential for resistance development in Bacillus cereus.
ATP synthase subunit c (atpE) shows distinctive conservation patterns across bacterial species, providing valuable insights into its evolution and functional importance:
Conservation Within Bacillales:
Taxonomic Distribution Patterns:
Variation increases with taxonomic distance from Bacillales
Core functional elements remain conserved across all bacteria
Transmembrane regions show higher conservation than connecting loops
The proton-binding carboxylate residue is nearly universally conserved
Structural Conservation vs. Sequence Divergence:
Despite sequence variations, the structural fold of atpE is highly conserved
This suggests strong structural constraints imposed by functional requirements
Conserved residues cluster at functionally critical sites (proton binding, subunit interfaces)
c-Ring Stoichiometry Variation:
The number of c-subunits forming the ring varies between species (typically 8-15)
This variation represents an evolutionary adaptation affecting the H⁺/ATP ratio
Reflects adaptation to different bioenergetic requirements across bacterial habitats
Selective Pressure Analysis:
This conservation pattern underscores the fundamental role of ATP synthase in bacterial bioenergetics while highlighting the evolutionary adaptations that allow specialization. The high conservation within Bacillales provides a rational basis for developing order-specific antimicrobial agents targeting atpE.
Comparative analysis of Bacillus cereus atpE with ATP synthase components in other pathogenic bacteria reveals important similarities and differences:
Structural Comparison with Other Gram-Positive Pathogens:
Differences from Gram-Negative Pathogens:
Greater sequence divergence compared to Proteobacteria (E. coli, Pseudomonas, etc.)
Structural differences in the c-ring, including potential variations in subunit number
Different lipid environment interactions affecting inhibitor access and binding
Mycobacterial ATP Synthase Comparison:
Mycobacterium tuberculosis ATP synthase has become an important drug target (bedaquiline)
B. cereus atpE shows significant structural differences from mycobacterial counterparts
These differences explain selective targeting of mycobacteria by bedaquiline
Provides insights for selective targeting of Bacillales
Functional Conservation Despite Sequence Variation:
Inhibitor Sensitivity Profiles:
This comparative understanding provides a foundation for developing pathogen-specific ATP synthase inhibitors with optimized selectivity profiles.
The most promising advances in ATP synthase research emerge from interdisciplinary approaches that integrate diverse scientific fields:
Structural Biology and Biochemistry Integration:
Combining high-resolution structural techniques (cryo-EM, X-ray crystallography) with functional assays
Correlating structural features with biochemical properties
Structure-guided mutagenesis to validate functional hypotheses
Example: Correlating ATP synthesis inhibition with structural binding sites of inhibitors
Chemical Biology and Medicinal Chemistry:
Rational design of ATP synthase inhibitors based on structural insights
Structure-activity relationship (SAR) studies to optimize inhibitor properties
Development of chemical probes to study ATP synthase function in vivo
As demonstrated with tomatidine derivatives that prevented high-level resistance development
Systems Biology and Bioenergetics:
Computational Biology and Biophysics:
Molecular dynamics simulations of c-ring rotation and proton translocation
Quantum mechanical modeling of proton transfer events
Machine learning approaches to predict inhibitor efficacy and resistance
In silico screening of compound libraries for novel inhibitors
Microbiology and Infectious Disease Research:
Linking ATP synthase function to bacterial pathogenesis
Understanding the role of energy metabolism in persistence and antibiotic tolerance
Translating basic ATP synthase research into therapeutic applications
Validating ATP synthase as a viable antimicrobial target in infection models
Evolutionary Biology and Comparative Genomics:
This interdisciplinary integration accelerates discovery by approaching ATP synthase from multiple perspectives, yielding insights that would be inaccessible through any single discipline. The success of identifying ATP synthase inhibitors as potential antibiotics demonstrates the power of this approach .