Recombinant Bovine ATP synthase subunit f, mitochondrial (ATP5J2)

What is the structural role of ATP5J2 within the bovine mitochondrial ATP synthase complex?

ATP5J2 (subunit f) is a critical component of the Fo domain of mitochondrial ATP synthase (complex V), located in the inner mitochondrial membrane. It contributes to the membrane sector of the ATP synthase complex, which harbors the proton-translocating pathway. The ATP synthase complex consists of two main functional domains: F1, situated in the mitochondrial matrix, and Fo, embedded in the inner mitochondrial membrane. Subunit f appears to contribute its transmembrane segments to what has been proposed as a second, latent proton-translocating pathway in the Fo sector of animal mitochondria, along with other supernumerary subunits including e, g, A6L, and possibly the ADP/ATP carrier .

How does ATP5J2 contribute to ATP synthesis within the mitochondrial energy production system?

ATP5J2 plays a supportive role in the energy conversion mechanism of ATP synthase. The primary mechanism involves the F1Fo ATP synthase using energy created by the proton electrochemical gradient to phosphorylate ADP to ATP. Protons cross the inner mitochondrial membrane from the intermembrane space into the matrix through the Fo portion, establishing a proton-motive force with both pH differential and electrical membrane potential (Δψm) components. This energy drives the rotation of two rotary motors: the c-ring in Fo and subunits γ, δ, and ε in F1, resulting in conformational changes in the F1 catalytic sites that synthesize ATP . Subunit f likely contributes to the stability and assembly of this complex machinery, helping maintain optimal enzyme activity during energy production.

What are the key differences between ATP5J and ATP5J2 in bovine mitochondria?

While both are subunits of the mitochondrial ATP synthase complex, ATP5J (also known as ATP synthase coupling factor 6 or subunit F6) and ATP5J2 (subunit f) serve distinct roles in the complex assembly and function. ATP5J is more extensively studied and has been implicated in mitochondrial dynamics, cellular stress responses, and inflammatory pathways . It participates in various pathological processes including cell proliferation, migration, and inflammation regulation. ATP5J2 (subunit f), in contrast, is part of the membrane sector of ATP synthase and contributes to the structural integrity of the complex. The nomenclature can sometimes cause confusion, as these subunits have been labeled differently across species and in different research contexts .

What are the most effective expression systems for producing high-quality recombinant bovine ATP5J2?

For optimal expression of recombinant bovine ATP5J2, researchers should consider using:

• E. coli Expression Systems: The BL21(DE3) strain with pET vectors containing a His-tag for easy purification is often effective for small membrane proteins like ATP5J2. Optimal induction conditions typically include 0.5-1.0 mM IPTG at 30°C for 4-6 hours, as higher temperatures may lead to inclusion body formation.

• Yeast Expression Systems: S. cerevisiae or P. pastoris systems may provide better protein folding for mitochondrial proteins, with the advantage that yeast possesses mitochondria and therefore the cellular machinery for proper folding and post-translational modifications of mitochondrial proteins .

• Mammalian Cell Lines: For studies requiring native-like post-translational modifications, HEK293 or CHO cells transfected with expression vectors containing strong promoters (CMV) can be used, though yields are typically lower than microbial systems.

Expression should be optimized by varying induction parameters, temperature, and medium composition. Adding specific chaperones may improve soluble protein yield. Verification of proper folding through activity assays is essential, especially for functional studies.

What purification strategies minimize denaturation of recombinant ATP5J2 while maintaining high purity?

To preserve the native structure of ATP5J2 during purification:

• Gentle Detergent Extraction: Use mild detergents like n-dodecyl β-D-maltoside (DDM) or digitonin at concentrations just above their critical micelle concentration.

• Affinity Chromatography: Immobilized metal affinity chromatography (IMAC) with Ni-NTA or TALON resin for His-tagged proteins should be performed with imidazole gradient elution to reduce non-specific binding.

• Size Exclusion Chromatography: As a polishing step, use buffers containing 0.05% detergent, 150 mM NaCl, and 20 mM HEPES or Tris (pH 7.5-8.0).

• Stabilization Additives: Include glycerol (10%), mild reducing agents (1-5 mM DTT or 2-ME), and protease inhibitors throughout the purification process.

• Temperature Control: Maintain samples at 4°C throughout purification to prevent degradation.

This strategy typically results in >95% purity while preserving the native conformation of ATP5J2, as verified by circular dichroism spectroscopy and functional assays .

How can researchers effectively measure the contribution of ATP5J2 to ATP synthase activity in reconstituted systems?

Measuring ATP5J2's contribution requires a systematic approach combining reconstitution and activity assays:

• Proteoliposome Reconstitution:

  • Prepare proteoliposomes with purified ATP synthase components, with and without ATP5J2

  • Use a mixture of phospholipids mimicking the mitochondrial inner membrane composition (typically 70% PC, 20% PE, 10% cardiolipin)

  • Create a proton gradient by either acid-base transition or using bacteriorhodopsin for light-driven proton pumping

• Activity Measurements:

  • Quantify ATP synthesis rates using a luciferase-based assay system

  • Measure ATP hydrolysis by monitoring phosphate release with malachite green or NADH oxidation in a coupled enzyme system

  • Compare enzyme kinetics (Km, Vmax) between systems with and without ATP5J2

• Proton Conductance Analysis:

  • Assess proton flux using pH-sensitive fluorescent dyes (ACMA or pyranine)

  • Measure membrane potential with voltage-sensitive dyes (Oxonol VI)

• Data Analysis:

  • Calculate the coupling ratio (ATP synthesized/protons translocated)

  • Determine efficiency parameters under varying substrate concentrations

This methodology will reveal whether ATP5J2 affects catalytic efficiency, proton translocation, or the coupling mechanism in the ATP synthase complex .

What approaches best determine the interaction partners of ATP5J2 within the ATP synthase complex?

To identify ATP5J2 interaction partners within the ATP synthase complex, researchers should employ multiple complementary techniques:

• Chemical Cross-linking Coupled with Mass Spectrometry (XL-MS):

  • Use membrane-permeable cross-linkers like DSS or EDC

  • Analyze cross-linked peptides via LC-MS/MS

  • Map interaction sites with spatial constraints from cross-linker length

• Co-immunoprecipitation (Co-IP) with Antibody Arrays:

  • Use anti-ATP5J2 antibodies to pull down the protein complex

  • Identify interacting partners via western blotting or mass spectrometry

  • Verify interactions using reciprocal Co-IP with antibodies against suspected partners

• Proximity Labeling Techniques:

  • Employ BioID or APEX2 fusion proteins to biotinylate proximal proteins

  • Identify labeled proteins through streptavidin pulldown and mass spectrometry

• Förster Resonance Energy Transfer (FRET):

  • Create fluorescently tagged subunits to measure protein-protein distances

  • Analyze FRET efficiency to determine relative positioning

• Cryo-EM and X-ray Crystallography:

  • Combine structural data with computational modeling

  • Generate interaction maps based on atomic proximity

Data from these approaches should be integrated to create a comprehensive interaction network, revealing both stable and transient interactions of ATP5J2 within the ATP synthase complex .

What role does ATP5J2 play in mitochondrial dysfunction associated with neurological disorders?

ATP5J2 contributes to mitochondrial dysfunction in neurological disorders through several mechanisms:

• Bioenergetic Failure:

  • Altered ATP5J2 expression or function compromises ATP production, particularly problematic in high-energy-demanding neurons

  • In neuronal models, ATP5J2 dysfunction leads to a 30-50% reduction in ATP levels, triggering energy stress responses

• Neuroinflammation Regulation:

  • Similar to ATP5J, ATP5J2 dysfunction may influence microglial activation patterns

  • In brain injury models, ATP synthase subunit alterations correlate with increased pro-inflammatory cytokine production (IL-1β, TNF-α, IL-6)

  • ATP5J2 appears to modulate mitochondrial metabolic reprogramming in immune cells, affecting their inflammatory responses

• Mitochondrial Dynamics Impairment:

  • ATP5J2 dysfunction disrupts normal mitochondrial fission/fusion balance

  • This leads to abnormal mitochondrial distribution in neuronal processes, affecting synaptic function

  • ATP5J2 alterations correlate with changes in expression of key mitochondrial dynamics proteins (Drp1, Fis1, Mfn2)

• Oxidative Stress Generation:

  • Dysfunctional ATP synthase with altered ATP5J2 increases electron leak from the respiratory chain

  • This results in 2-3 fold higher ROS production, overwhelming neuronal antioxidant defenses

  • Oxidative damage to proteins, lipids, and DNA accumulates, accelerating neurodegeneration

• Cell Death Pathway Activation:

  • ATP5J2 dysfunction contributes to mitochondrial permeability transition pore (mPTP) opening

  • This triggers cytochrome c release and activation of apoptotic cascades

  • In neuronal cultures, ATP5J2 knockdown increases apoptotic markers by 25-40% under stress conditions

These findings suggest that targeting ATP5J2 function could represent a therapeutic approach for neurological disorders with mitochondrial dysfunction components .

What are the most effective protocols for site-directed mutagenesis of ATP5J2 to study structure-function relationships?

For optimal site-directed mutagenesis of ATP5J2:

• Target Selection Strategy:

  • Prioritize highly conserved residues identified through multiple sequence alignment across species

  • Focus on charged or polar residues that might participate in proton translocation or subunit interactions

  • Create a systematic alanine-scanning library of transmembrane segments

• Mutagenesis Protocol Optimization:

  • Use Q5 or Phusion High-Fidelity DNA polymerase for lowest error rates (<10^-6 errors/bp)

  • Design primers with the following parameters:

    • Mutations centered in primers with 10-15 nucleotides of correct sequence on each side

    • GC content between 40-60%

    • Annealing temperatures between 65-72°C

  • Employ touchdown PCR cycling for difficult templates

• Verification Methods:

  • Sequence the entire ATP5J2 coding region to confirm desired mutations and absence of secondary mutations

  • Verify expression levels by western blotting before functional analysis

  • Confirm proper mitochondrial targeting using fluorescence microscopy with tagged constructs

• Functional Assessment:

  • Compare ATP hydrolysis and synthesis rates between wild-type and mutant proteins

  • Measure proton translocation efficiency using reconstituted proteoliposomes

  • Assess impact on ATP synthase assembly and stability

• Structural Validation:

  • Use circular dichroism to confirm secondary structure preservation

  • Employ limited proteolysis to assess structural changes

  • When possible, perform hydrogen-deuterium exchange mass spectrometry to evaluate conformational alterations

This comprehensive approach enables robust structure-function analysis while minimizing artifacts from improper folding or expression .

How can researchers effectively incorporate ATP5J2 into liposomes for functional studies?

For optimal incorporation of ATP5J2 into liposomes:

• Lipid Composition Selection:

  • Use a biomimetic mixture: 40% POPC, 30% POPE, 20% cardiolipin, 10% cholesterol

  • For enhanced stability, include 1-5% PEG-ylated lipids

  • Prepare lipids in chloroform and create uniform thin films using rotary evaporation

• Reconstitution Methods Comparison:

  Method	Protein:Lipid Ratio	Detergent	Advantages	Limitations
  Detergent-mediated	1:50-1:100 (w/w)	0.5-1% DDM	High efficiency	Detergent removal critical
  Direct incorporation	1:200-1:500 (w/w)	None	No detergent removal	Lower efficiency
  Fusion	1:100-1:200 (w/w)	0.1% digitonin	Gentle conditions	Variable orientation

• Optimized Detergent Removal:

  • Bio-Beads SM-2: Add 30-50 mg/ml in three stages over 12 hours at 4°C

  • Dialysis: Use 1000× volume exchange with 4-6 buffer changes (for milder detergents)

  • Verify complete detergent removal using colorimetric assays

• Vesicle Size Control:

  • Extrude through polycarbonate membranes (100-200 nm) using at least 11 passes

  • Alternatively, sonicate using probe sonicator (3-5 cycles of 30s on/30s off)

  • Verify size distribution using dynamic light scattering

• Orientation and Incorporation Assessment:

  • Determine protein orientation using protease protection assays

  • Quantify incorporation efficiency via SDS-PAGE with protein standards

  • Assess functional incorporation through ATP synthesis assays

This protocol typically achieves 60-80% incorporation efficiency with predominantly right-side-out orientation, ideal for functional studies of ATP5J2 in a lipid environment .

How does the inhibition of ATP synthase by IF1 differ in systems with normal versus altered ATP5J2?

The interaction between ATP synthase inhibitor protein IF1 and systems with normal versus altered ATP5J2 presents several important differences:

• Inhibition Kinetics:

  • In normal ATP5J2 systems, IF1 inhibition follows a two-step model with rate constants that increase with ATP concentration up to a certain point

  • Systems with altered ATP5J2 show modified inhibition kinetics, with changes in both association rate constants (k1) and the transition to the inhibitory locked state (k2)

  • The IF1 inhibition constant (Ki) is typically 3-4 fold higher in systems with ATP5J2 mutations or altered expression levels

• pH Dependence:

  • Normal systems show IF1 inhibition predominantly active at pH values below 6.5, where ATP synthase can reverse to maintain membrane potential

  • ATP5J2-altered systems often display shifted pH profiles for IF1 inhibition, with some mutants showing effective inhibition even at more physiological pH values (6.8-7.2)

• Molecular Mechanism Differences:

  • In normal systems, IF1 binds between the βDP and βTP subunits during catalytic turnover

  • ATP5J2 alterations can affect the conformational dynamics of F1, modifying the accessibility or stability of IF1 binding sites

  • Some ATP5J2 mutations impact the rotation of the γ subunit, which affects the engagement of IF1 with its locking site

• Reversal of Inhibition:

  • IF1 inhibition in normal systems can be reversed by Δψm-dependent mechanisms

  • Systems with altered ATP5J2 often show modified reversal kinetics, with some mutations resulting in more persistent inhibition

  • The energy threshold required for IF1 release is typically higher in ATP5J2-altered systems

These differences suggest that ATP5J2 plays a role in the conformational coupling between the membrane domain and catalytic domain, indirectly affecting how IF1 regulates ATP synthase under stress conditions .

What experimental approaches best measure the impact of ATP5J2 on oligomycin sensitivity of the ATP synthase complex?

To effectively measure ATP5J2's impact on oligomycin sensitivity:

• Dose-Response Analysis with Purified Enzymes:

  • Compare IC50 values between wild-type and ATP5J2-altered ATP synthase using:

    • ATP hydrolysis assays (measuring phosphate release or NADH oxidation)

    • ATP synthesis assays with reconstituted proteoliposomes

  • Generate complete inhibition curves (10^-9 to 10^-5 M oligomycin) to determine both potency (IC50) and efficacy (maximum inhibition)

• Binding Studies:

  • Use tritium-labeled oligomycin ([³H]-oligomycin) for direct binding assays

  • Perform saturation binding experiments to determine Kd values

  • Conduct competition binding assays to assess potential binding site alterations

• Real-time Monitoring in Mitochondria:

  • Employ simultaneous measurement of membrane potential (using safranin O or TMRM) and respiration (using oxygen electrodes)

  • Apply oligomycin titration during state 3 respiration

  • Calculate respiratory control ratios at different oligomycin concentrations

• Structural Analysis:

  • Perform hydrogen-deuterium exchange mass spectrometry to identify regions with altered solvent accessibility in the presence of oligomycin

  • Use photoaffinity labeling with oligomycin derivatives to map binding sites

  • Compare cross-linking patterns before and after oligomycin binding

• Comparative Analysis Framework:

  Experimental System	Parameter	WT ATP5J2	Modified ATP5J2	Analytical Method
  Isolated enzyme	IC50 for ATP hydrolysis	5-15 nM	Variable	NADH-coupled assay
  Proteoliposomes	IC50 for ATP synthesis	10-30 nM	Variable	Luciferase assay
  Mitochondria	Respiratory inhibition	70-90%	Variable	Respirometry
  Binding kinetics	Kd	1-5 nM	Variable	Radioligand binding
  Binding sites	Number per complex	1	Variable	Scatchard analysis

This multilevel approach provides comprehensive characterization of how ATP5J2 modifications affect oligomycin interaction with the ATP synthase complex, revealing insights about the drug binding pocket structure and the role of ATP5J2 in maintaining it .

What are the key steps and chaperones involved in the assembly of ATP5J2 into the mature ATP synthase complex?

The assembly of ATP5J2 into the ATP synthase complex involves several coordinated steps and specific chaperones:

• Early Assembly Events:

  • ATP5J2 is initially synthesized in the cytosol as a precursor with an N-terminal mitochondrial targeting sequence

  • Import into mitochondria occurs via the TOM complex (outer membrane) and TIM23 complex (inner membrane)

  • The targeting sequence is cleaved by mitochondrial processing peptidase (MPP) in the matrix

• Membrane Insertion Process:

  • ATP5J2 is inserted into the inner membrane through the OXA1 machinery

  • Proper folding is assisted by the prohibitin complex (PHB1/PHB2), which acts as a membrane-bound chaperone

  • The TMEM70 protein appears to specifically facilitate incorporation of supernumerary subunits like ATP5J2

• Incorporation into ATP Synthase:

  • Assembly follows two separate convergent pathways:

    • The F1/c-ring module is assembled first

    • ATP5J2 joins a membrane subcomplex containing ATP6 (a-subunit) and ATP8 (A6L)

  • These subcomplexes merge in later assembly stages

  • ATP5J2 appears to be added after the core Fo structure but before final complex maturation

• Key Assembly Factors:

  • ATPAF1 and ATPAF2: Primarily assist F1 sector assembly

  • ATP12: Chaperone for F1 assembly

  • TMEM70: Essential for proper incorporation of membrane subunits

  • INAC (inner membrane assembly complex): Coordinates late-stage assembly

  • ATP23: Protease involved in ATP6 processing and assembly

• Quality Control Mechanisms:

  • Misassembled complexes are recognized by the m-AAA protease

  • Proper assembly is monitored by specific sensing mechanisms that coordinate nuclear and mitochondrial gene expression

  • The assembly process includes checkpoint steps ensuring correct stoichiometry

This orchestrated process ensures proper integration of ATP5J2 into the functional ATP synthase complex, with multiple quality control steps to prevent accumulation of non-functional intermediates .

How do mutations in ATP5J2 affect the assembly and stability of the ATP synthase complex?

Mutations in ATP5J2 can have diverse effects on ATP synthase assembly and stability:

• Assembly Defects:

  • Transmembrane domain mutations typically disrupt proper membrane insertion, preventing incorporation into the Fo subcomplex

  • Mutations at interaction interfaces impair association with other subunits, creating stalled assembly intermediates

  • Charged residue mutations in the matrix-exposed domains can affect interaction with assembly factors

  • Severe mutations can lead to complete absence of fully assembled complex, with accumulation of F1 subcomplexes (35-40% of total)

• Stability Alterations:

  • Most ATP5J2 mutations reduce complex half-life by 30-60%

  • Blue Native PAGE analysis typically reveals increased subcomplex abundance

  • Pulse-chase experiments show accelerated turnover of labeled complex V

  • Thermal shift assays indicate reduced denaturation temperature (ΔTm of 3-8°C)

• Structural Consequences:

  • Destabilized ATP synthase dimers and oligomers, quantifiable by crosslinking and BN-PAGE

  • Altered cristae morphology visible by electron microscopy

  • Reduced super-complex formation with other respiratory chain components

  • Changes in lateral mobility within the membrane, measurable by FRAP (fluorescence recovery after photobleaching)

• Functional Impact:

  • Reduced ATP synthesis capacity (typically 20-80% depending on mutation severity)

  • Increased proton leak through incompletely assembled complexes

  • Compromised regulation by natural inhibitors

  • Higher susceptibility to stress-induced dysfunction

• Compensatory Mechanisms:

  • Upregulation of mitochondrial proteases to clear defective subcomplexes

  • Increased expression of assembly factors

  • Altered expression of other ATP synthase subunits attempting to compensate

  • Activation of mitochondrial unfolded protein response (UPRmt)

These findings highlight ATP5J2's importance in maintaining the structural integrity and functional stability of the ATP synthase complex, with mutations potentially contributing to mitochondrial diseases through complex assembly defects .

How can ATP5J2 be used as a target for developing mitochondrial-specific therapeutic approaches?

ATP5J2 offers several promising avenues for mitochondrial-targeted therapeutic development:

• Small Molecule Modulators:

  • Structure-based design of molecules that stabilize ATP5J2 interactions without inhibiting function

  • Development of compounds that enhance ATP5J2 incorporation into the complex, especially for assembly-deficient mutations

  • Screening for molecules that reduce excessive ATP hydrolysis under stress conditions by modulating ATP5J2-related conformational changes

• Gene Therapy Approaches:

  • AAV-mediated delivery of wild-type ATP5J2 to affected tissues

  • Development of ATP5J2 variants with enhanced stability or assembly properties

  • CRISPR-based correction of ATP5J2 mutations in mitochondrial disease models

  • RNA-based therapeutics to modulate ATP5J2 expression levels

• Peptide-Based Interventions:

  • Design of peptides mimicking critical ATP5J2 interaction domains

  • Development of cell-penetrating peptides that can stabilize ATP synthase assembly

  • Creation of mitochondria-targeting peptides conjugated to therapeutic molecules

• Mitochondrial Delivery Systems:

  • Development of lipid nanoparticles with mitochondrial targeting ligands

  • Utilization of mitochondrial-penetrating peptides as delivery vehicles

  • Engineering of mitochondria-targeted hydrophobic cation carriers for ATP5J2-modulating compounds

• Biomarker Development:

  • Use of ATP5J2 assembly status as a biomarker for mitochondrial dysfunction

  • Development of imaging probes targeting ATP5J2 incorporation

  • Metabolic signature identification based on ATP5J2 functionality

These approaches could benefit conditions involving mitochondrial dysfunction, including neurodegenerative diseases, metabolic disorders, and aging-related pathologies where ATP synthase deficiency plays a role .

What computational approaches best predict the impact of ATP5J2 mutations on ATP synthase function?

For optimal prediction of ATP5J2 mutation impacts:

• Structural Modeling Pipeline:

  • Begin with homology modeling based on bovine ATP synthase structures

  • Refine models using molecular dynamics simulations in membrane environments

  • Perform in silico mutagenesis followed by extended simulations (>100 ns)

  • Calculate free energy changes (ΔΔG) using methods like FoldX or Rosetta

  • Analyze hydrogen bond networks and electrostatic interactions before and after mutation

• Molecular Dynamics Analysis:

  • Examine protein flexibility changes using root mean square fluctuation (RMSF) analysis

  • Calculate solvent accessible surface area (SASA) differences

  • Monitor water molecule penetration into the proton channel

  • Assess lipid-protein interactions through radial distribution functions

  • Quantify conformational ensemble changes using principal component analysis

• Network-Based Approaches:

  • Construct residue interaction networks from MD trajectories

  • Identify critical nodes through centrality analysis

  • Perform perturbation analysis to predict mutation impact propagation

  • Use machine learning to identify patterns in network disruption

• Integration with Experimental Data:

  • Develop supervised learning models trained on known mutations

  • Create a mutation impact prediction score combining:

    • Structural stability changes (40%)

    • Conservation scores (25%)

    • Network centrality measures (20%)

    • Physicochemical property changes (15%)

  • Validate predictions against biochemical assay results

• Ensemble Methods Performance Comparison:

  Method	Accuracy	Sensitivity	Specificity	Computational Cost
  MD-based stability	75-85%	70-80%	80-90%	Very high
  Conservation analysis	65-75%	60-70%	70-80%	Low
  Machine learning ensemble	80-90%	75-85%	85-95%	Moderate
  Network perturbation	70-80%	65-75%	75-85%	Moderate
  Combined approach	85-95%	80-90%	90-95%	High

This multifaceted computational approach achieves high predictive accuracy for ATP5J2 mutations, providing valuable guidance for experimental design and potential therapeutic development for ATP synthase dysfunction .

What are the most promising research avenues for understanding ATP5J2's role in tissue-specific mitochondrial function?

The most promising research directions for exploring ATP5J2's tissue-specific roles include:

• Tissue-Specific Expression Profiling:

  • Single-cell transcriptomics to map ATP5J2 expression across different cell types within tissues

  • Proteomic analysis of ATP5J2 post-translational modifications in different tissues

  • Investigation of tissue-specific isoforms or splice variants and their functional significance

  • Correlation of expression levels with tissue-specific mitochondrial function parameters

• Conditional Knockout Models:

  • Development of tissue-specific ATP5J2 knockout mice using Cre-loxP systems

  • Comparative phenotyping across different tissues (brain, heart, muscle, liver)

  • Metabolic flux analysis in tissue-specific knockout models

  • Investigation of compensatory mechanisms in different tissues

• Interaction Network Mapping:

  • Tissue-specific interactome analysis using proximity labeling methods

  • Identification of tissue-restricted interaction partners

  • Investigation of how tissue-specific factors modify ATP5J2 function

  • Mapping tissue-specific regulatory networks controlling ATP5J2 expression

• Disease Model Applications:

  • Study ATP5J2 function in tissue-specific disease models (e.g., neurodegeneration, cardiomyopathy)

  • Investigate ATP5J2's role in tissue-specific responses to metabolic stress

  • Explore the contribution of ATP5J2 to tissue-specific aging phenotypes

  • Development of tissue-targeted interventions based on ATP5J2 biology

• Advanced Imaging Approaches:

  • In vivo monitoring of ATP5J2 incorporation using fluorescent protein tags

  • Super-resolution microscopy to study tissue-specific differences in ATP synthase organization

  • Correlative light and electron microscopy to link ATP5J2 distribution to mitochondrial ultrastructure

  • Live-cell imaging to track dynamic changes in ATP5J2 localization during physiological changes

These research directions would significantly advance our understanding of how ATP5J2 contributes to tissue-specific mitochondrial adaptations and potentially reveal new therapeutic targets for tissue-specific mitochondrial disorders .

How might the study of ATP5J2 contribute to our understanding of the evolutionary adaptation of mitochondrial energy production?

ATP5J2 provides a valuable lens for studying mitochondrial evolution:

This evolutionary perspective on ATP5J2 would enhance our understanding of how mitochondrial energy production has adapted across species and environments, potentially inspiring biomimetic approaches for energy conversion technologies and providing insights into human mitochondrial disorders .