Recombinant Arabidopsis thaliana NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13-B (At2g33220)

What is the molecular structure and function of At2g33220?

At2g33220 (NADH-ubiquinone oxidoreductase B 16.6 kDa subunit) is a nuclear-encoded mitochondrial protein in Arabidopsis thaliana with a molecular weight of approximately 16.0 kD as determined from nucleotide sequence analysis, which aligns with the 16 kD experimental measurement . The protein functions as one of the multiple subunits that constitute the NADH-ubiquinone oxidoreductase (Complex I) in the mitochondrial electron transport chain.

This particular subunit contributes to the structural integrity of Complex I and participates in the electron transfer process from NADH to ubiquinone. Complex I catalyzes the first step of the mitochondrial respiratory chain, oxidizing NADH derived from metabolic pathways such as the TCA cycle, pyruvate dehydrogenase activity, and glycine oxidation in green leaf mitochondria . As part of this process, At2g33220 assists in the coupling of electron transfer to proton translocation across the inner mitochondrial membrane, which is essential for generating the proton motive force required for ATP synthesis.

The protein contains characteristic domains identified through database annotations, including membership in protein families as indicated by InterPro (IPR009346) and Pfam (PF06212) classifications . These conserved domains suggest evolutionary preservation of function across species and highlight the fundamental importance of this subunit in mitochondrial respiration.

How does At2g33220 integrate into the respiratory complex I structure?

At2g33220 functions as a specific subunit (16.6 kDa B subunit) within the larger NADH-ubiquinone oxidoreductase complex in Arabidopsis thaliana. The complete complex consists of at least 30 distinct protein subunits that work together to form the functional respiratory Complex I. At2g33220 is part of a specific assembly module within this larger structure.

The integration of At2g33220 occurs during the stepwise assembly process of Complex I, where various subcomplexes form intermediate structures before final assembly. Based on current understanding, the protein likely contributes to the peripheral arm of Complex I that extends into the mitochondrial matrix, where NADH oxidation occurs. The specific positioning of At2g33220 is crucial for maintaining proper electron flow from NADH through the iron-sulfur clusters to ubiquinone.

The complex organization can be visualized in the following subunit composition table:

This table demonstrates the diverse composition of Complex I and places At2g33220 within the context of the complete respiratory complex structure .

What experimental methods are most effective for studying At2g33220 expression and localization?

Several experimental approaches have proven effective for investigating At2g33220 expression patterns and subcellular localization:

• Transcriptomic Analysis: RNA-Seq and microarray techniques can quantify At2g33220 transcript levels across different tissues, developmental stages, and stress conditions. Cross-referencing with databases like ArrayExpress (O49313) can provide expression profiles across various experimental conditions .

• Protein Isolation and Western Blotting: Microsomal fractions can be isolated through differential centrifugation followed by protein separation using SDS-PAGE. Western blot analysis using antibodies specific to At2g33220 allows quantification of protein expression levels. This approach was effectively utilized in studies analyzing mitochondrial protein changes under microgravity conditions .

• Mass Spectrometry: LC-MS/MS analysis of mitochondrial fractions provides comprehensive proteome characterization. An effective workflow involves:

  • Sample preparation by reduction, alkylation, and trypsin digestion

  • Peptide separation using C18 columns

  • Mass spectrometry using an LTQ-Orbitrap system

  • Data validation through comparison with protein databases

• Fluorescence Microscopy with Tagged Constructs: GFP or other fluorescent protein fusions with At2g33220 can visualize subcellular localization in living cells. This approach can confirm mitochondrial targeting and potential dynamic changes under different conditions.

• Mitochondrial Isolation and Functional Assays: Isolating intact mitochondria allows measurement of NADH oxidation rates, which can indirectly assess At2g33220 function within Complex I. Measurements of oxygen consumption using Clark-type electrodes provide functional data on respiratory complex activity .

• T-DNA Insertion Lines and Transgenic Approaches: Functional studies using knockout or overexpression lines provide insights into phenotypic consequences of altered At2g33220 expression. Similar approaches with related proteins have demonstrated their utility in understanding functional roles in stress responses .

Each of these methodologies offers distinct advantages depending on the specific research question being addressed. For comprehensive characterization, combining multiple approaches provides the most robust results.

How does At2g33220 contribute to electron transport mechanisms compared to other complex I subunits?

At2g33220 functions as the NADH-ubiquinone oxidoreductase B 16.6 kDa subunit within Complex I, and its specific contribution to electron transport must be considered in relation to other subunits. While direct experimental data on At2g33220's precise electron transport role is limited in the provided search results, we can infer its function based on studies of related subunits and Complex I structure-function relationships.

• Stabilizing protein-protein interactions within the complex

• Modulating conformational changes during electron transfer

• Influencing the redox properties of nearby electron carriers

• Providing binding sites for regulatory molecules

What are the challenges in expressing and purifying recombinant At2g33220 for structural studies?

Expressing and purifying recombinant At2g33220 presents several significant challenges that researchers must address for successful structural studies:

• Membrane Protein Expression Issues: As a component of the membrane-bound Complex I, At2g33220 contains hydrophobic regions that can cause aggregation or misfolding when expressed recombinantly. Expression systems must be carefully optimized to maintain proper folding.

• Mitochondrial Targeting Sequence Handling: The protein contains mitochondrial targeting sequences that direct it to mitochondria in vivo. For recombinant expression, decisions must be made whether to include or exclude these sequences, which can affect solubility and folding.

• Requirement for Detergents or Membrane Mimetics: Purification typically requires detergents or membrane mimetics to maintain protein stability. The choice of detergent is critical and may need extensive optimization to preserve native structure without interfering with downstream applications.

• Co-expression Requirements: At2g33220 may require co-expression with interacting partners for proper folding and stability. As seen in the search results, At2g33220 functions as part of a multi-subunit complex , and isolation of individual subunits may result in instability.

• Post-translational Modifications: Any post-translational modifications present in the native protein may be absent in recombinant systems, potentially affecting structure and function.

The methods section from the microgravity experiment provides a useful protocol that could be adapted for recombinant protein work:

• Use of microsomal fractions

• Careful denaturation and reduction with Laemmli buffer at 95°C

• Alkylation with 90 mM iodoacetamide

• Protein concentration determination using detergent-compatible assays

• Appropriate buffer selection for maintaining stability

For structural studies specifically, X-ray crystallography or cryo-electron microscopy would require highly pure, homogeneous, and stable protein preparations. Given these challenges, alternative approaches such as expressing smaller domains or creating fusion proteins with solubility-enhancing tags might be necessary to facilitate structural characterization of At2g33220.

How does environmental stress affect At2g33220 expression and function?

Environmental stress significantly impacts At2g33220 expression and function as part of the plant's adaptive response mechanisms. While the search results don't provide direct data on At2g33220's stress response specifically, we can draw insights from studies on related components of the plant respiratory apparatus.

Research on alternative NAD(P)H dehydrogenases and oxidases in Arabidopsis demonstrates that components of the mitochondrial electron transport chain respond dynamically to environmental stressors. Plants lacking either AtAOX1A or AtNDB2 (another NAD(P)H dehydrogenase component) were more sensitive to combined drought and elevated light treatments, whereas plants overexpressing these components showed increased tolerance and capacity for post-stress recovery .

This suggests that At2g33220, as part of Complex I, likely participates in adaptive responses to environmental stress through:

• Altered Expression Patterns: Transcriptional or translational regulation may increase or decrease At2g33220 levels depending on the specific stress and the plant's energy demands.

• Modified Protein Interactions: Stress conditions may alter At2g33220's interactions with other Complex I subunits or regulatory proteins, potentially redirecting electron flow through alternative pathways.

• Contribution to Redox Homeostasis: As part of the electron transport system, At2g33220 may help maintain cellular redox balance during stress conditions when ROS production increases.

• Response to Microgravity: The GENARA A experiment studied how Arabidopsis proteome changes under microgravity conditions, which is an extreme environmental stress. The experimental procedures described in search result outline methods for detecting protein-level changes in response to this stress, which could be applied to study At2g33220 specifically.

Experimental approaches to further investigate At2g33220's role in stress response could include:

• qRT-PCR analysis of transcript levels under various stress conditions

• Proteomics analysis of mitochondrial fractions from stressed plants

• Functional analysis of electron transport in isolated mitochondria exposed to stress conditions

• Phenotypic analysis of At2g33220 knockout or overexpression lines under stress challenges

These approaches would help clarify whether At2g33220 functions primarily in maintaining basic respiratory capacity during stress or plays a more specific regulatory role in stress adaptation.

What are the optimal conditions for isolating functional mitochondria to study At2g33220 activity?

Isolating functional mitochondria with preserved At2g33220 activity requires careful attention to detail at each step of the procedure. Based on established protocols for plant mitochondrial isolation, the following conditions are optimal:

• Plant Material Selection and Preparation:

  • Use young, actively growing tissue (7-14 day old seedlings are optimal)

  • Harvest material at a consistent time of day to minimize circadian variations

  • Immediately process tissue or flash-freeze in liquid nitrogen if immediate processing is not possible

• Homogenization Buffer Composition:

  • 0.3 M sucrose (to maintain osmotic balance)

  • 25 mM MOPS-KOH, pH 7.5 (for pH buffering)

  • 1 mM EGTA (to chelate calcium)

  • 0.1% BSA (to scavenge fatty acids)

  • 0.6% PVPP (to remove phenolics)

  • 1 mM DTT (to maintain reducing environment)

  • Protease inhibitor cocktail (to prevent protein degradation)

• Temperature Control:

  • Maintain all solutions and equipment at 4°C

  • Perform all steps on ice or in cold room

  • Pre-chill all centrifuges to 4°C

• Isolation Procedure:

  • Gentle homogenization to avoid damaging mitochondria

  • Differential centrifugation series (typically 1,000 × g to remove debris, followed by 12,000 × g to pellet mitochondria)

  • Density gradient purification using Percoll gradients (18-23-40%)

• Functional Assessment:

  • Measure oxygen consumption rates using a Clark-type electrode

  • Assess NADH oxidation rates using spectrophotometric methods similar to those used in the AtNDB2 studies

  • Verify membrane integrity using cytochrome c test

For specific measurement of At2g33220 activity within Complex I, the following assay conditions have proven effective:

• Reaction buffer containing 0.3 M sucrose, 10 mM TES-KOH (pH 7.5), and 5 mM KH₂PO₄

• NADH concentration of 1-2 mM

• Temperature maintained at 25°C

• Inhibitors like rotenone can be used to distinguish Complex I activity from alternative dehydrogenases

• Activity measured as the decrease in absorbance at 340 nm as NADH is oxidized

The search results indicate that measured NADH oxidation rates in wild-type Arabidopsis mitochondria typically range from 20-30 nmol NADH min⁻¹ mg⁻¹ protein , providing a benchmark for comparison when assessing the integrity of isolated mitochondria and At2g33220 function.

How can transgenic approaches be used to study At2g33220 function in planta?

Transgenic approaches offer powerful tools for investigating At2g33220 function within the living plant. Based on successful strategies employed for similar mitochondrial proteins, the following approaches are most effective:

• T-DNA Insertion Lines:

  • T-DNA insertion mutants targeting At2g33220 can provide loss-of-function phenotypes

  • When working with T-DNA lines, it's crucial to:

    • Confirm insertion location through PCR and sequencing

    • Verify protein absence using Western blotting

    • Assess mitochondrial function through respiratory measurements

    • This approach revealed that disruption of AtNDB2 led to a 90% decrease in external NADH oxidation in isolated mitochondria

• RNAi or CRISPR-Cas9 for Gene Silencing/Knockout:

  • When T-DNA lines are unavailable or insertions are lethal, RNAi or CRISPR approaches provide alternatives

  • Design target-specific constructs to minimize off-target effects

  • Use inducible or tissue-specific promoters if constitutive knockdown causes lethality

• Overexpression Systems:

  • Constitutive overexpression using the CaMV 35S promoter

  • Inducible overexpression systems (e.g., estradiol-inducible) for controlled expression

  • Tissue-specific overexpression to study function in particular plant organs

  • Research shows that overexpression of related proteins like AtNDB2 led to increased protein abundance in mitochondria, though functional enhancement may require coordinated expression with interacting partners

• Fluorescent Protein Fusions:

  • C- or N-terminal GFP fusions for subcellular localization studies

  • Split-GFP approaches for detecting protein-protein interactions

  • FRET-based systems for studying dynamic interactions with other Complex I subunits

• Complementation Studies:

  • Express At2g33220 in knockout backgrounds to confirm phenotypic rescue

  • Express modified versions (point mutations, truncations) to identify functional domains

  • Cross-species complementation to assess functional conservation

• Stress Response Analysis:

  • Subject transgenic lines to environmental stressors (drought, high light, temperature extremes)

  • Monitor stress tolerance parameters (growth, photosynthetic efficiency, ROS production)

  • Compare recovery capacity following stress exposure

  • Studies of related proteins showed that plants lacking either AtAOX1A or AtNDB2 were more sensitive to combined drought and elevated light treatments

The search results demonstrate that these approaches have successfully revealed functional aspects of related mitochondrial proteins. For example, overexpression of AtNDB2 alone did not significantly enhance external NADH oxidation unless AtAOX1A was concomitantly overexpressed and activated, revealing important functional interactions between mitochondrial components .

What mass spectrometry protocols are most effective for At2g33220 identification and quantification?

Effective mass spectrometry protocols for At2g33220 identification and quantification should be tailored to address the specific challenges of analyzing mitochondrial membrane proteins. Based on the search results and established practices in the field, the following comprehensive protocol is recommended:

• Sample Preparation:

  • Mitochondrial Isolation: Isolate mitochondria using differential centrifugation followed by Percoll gradient purification

  • Membrane Protein Extraction: Use a combination of detergents (e.g., 1% digitonin or 0.5% n-dodecyl-β-D-maltoside) to solubilize membrane proteins

  • Protein Denaturation: Heat samples at 95°C for 30 minutes in Laemmli buffer containing appropriate reducing agents

  • Alkylation: Treat with 90 mM iodoacetamide for 30 minutes at room temperature in the dark to prevent disulfide bond reformation

  • Concentration Determination: Use detergent-compatible protein assays (e.g., Bio-Rad Protein Assay Kit) to determine protein concentration

• Protein Separation:

  • Gel-based Approach: Concentrate proteins in a single band on 12% acrylamide SDS-PAGE gel and visualize with colloidal Coomassie Blue staining

  • Alternative Approach: Blue native PAGE can separate intact complexes while maintaining native structure, allowing identification of At2g33220 within Complex I

• In-gel Digestion:

  • Washing: Wash gel pieces in 50 mM ammonium bicarbonate (15 min, 37°C) followed by 50 mM ammonium bicarbonate/acetonitrile (1:1, 15 min, 37°C)

  • Trypsin Digestion: Perform overnight digestion at 37°C with high-quality trypsin (e.g., Promega)

  • Peptide Extraction: Extract peptides using sequential incubations in 50 mM ammonium bicarbonate (15 min, 37°C) and two incubations in 10% formic acid/acetonitrile (1:1, 15 min, 37°C)

• LC-MS/MS Analysis:

  • Nano-LC System: Ultimate3000 system (Dionex) or equivalent

  • Mass Spectrometer: High-resolution instrument such as LTQ-Orbitrap Velos (Thermo Fisher Scientific)

  • Sample Loading: Load peptides on a C18 precolumn (300 μm inner diameter × 5 mm) at 20 μl/min in 5% acetonitrile, 0.05% trifluoroacetic acid

  • Desalting: 5 minutes on precolumn

  • Analytical Separation: C18 column (75-μm inner diameter × 15 cm) with gradient of 5-50% solvent B (80% acetonitrile, 0.2% formic acid) over 110 minutes at 300 nl/min

• Data Analysis and Quantification:

  • Database Searching: Search against Arabidopsis thaliana protein database with appropriate parameters (e.g., variable modifications for oxidation, fixed modifications for carbamidomethylation)

  • Identification Criteria: Minimum of two unique peptides with false discovery rate < 1%

  • Quantification Methods:

    • Label-free quantification based on spectral counting or intensity

    • Isotope labeling approaches (SILAC, TMT, iTRAQ) for more accurate quantification

    • Selected reaction monitoring (SRM) for targeted quantification of specific At2g33220 peptides

This protocol, adapted from the GENARA A experiment methodology , provides a robust framework for reliable identification and quantification of At2g33220 in complex mitochondrial samples. The high sensitivity of modern mass spectrometry systems allows detection of low-abundance membrane proteins like At2g33220, while appropriate sample preparation methods ensure maximum recovery and identification of representative peptides.

How does At2g33220 contribute to plant stress responses and adaptation mechanisms?

At2g33220, as a component of mitochondrial Complex I, likely plays a significant role in plant stress responses and adaptation mechanisms. While the search results don't specifically address At2g33220's role in stress, we can extrapolate from studies on related components of the respiratory apparatus to understand its potential contributions.

Several mechanisms likely involve At2g33220 in stress adaptation:

• Maintenance of Energy Production During Stress:

  • Complex I generates a significant portion of the proton motive force needed for ATP synthesis

  • During stress, maintaining ATP production is crucial for powering defense mechanisms and repair processes

  • At2g33220, as a structural/functional component of Complex I, contributes to this energy production capacity

• Regulation of Cellular Redox Balance:

  • Complex I oxidizes NADH to NAD+, affecting cellular NADH/NAD+ ratios

  • This redox balance is crucial during stress when metabolic adjustments must occur rapidly

  • Proper functioning of At2g33220 within Complex I ensures efficient NADH oxidation

• Coordination with Alternative Respiratory Pathways:

  • Studies show that components of the alternative pathway (AtNDB2 and AtAOX1A) enhance stress tolerance

  • Plants lacking these components showed increased sensitivity to combined drought and elevated light treatments

  • As part of the main respiratory chain, At2g33220 likely works in concert with these alternative pathways to optimize respiration under stress

• Potential Stress-Specific Regulation:

  • Expression or post-translational modifications of At2g33220 may change under stress conditions

  • Such changes could alter Complex I activity or assembly, influencing respiratory efficiency

  • Microgravity experiments demonstrated that environmental stress can induce changes in the proteome of Arabidopsis membranes

• Contribution to Recovery Mechanisms:

  • Plants overexpressing components of the alternative respiratory pathway showed increased capacity for post-stress recovery

  • At2g33220 may similarly contribute to recovery processes by ensuring rapid restoration of respiratory capacity after stress

Experimental approaches to further elucidate At2g33220's role in stress response could include comparative analysis of wild-type and At2g33220 mutant plants under various stress conditions, similar to studies performed with AtNDB2 and AtAOX1A mutants. These studies could assess parameters such as growth, photosynthetic efficiency, respiratory rates, ROS production, and metabolite profiles to build a comprehensive picture of At2g33220's contribution to stress adaptation.

What protein-protein interactions does At2g33220 participate in within Complex I and potentially with other mitochondrial components?

At2g33220 (NADH-ubiquinone oxidoreductase B 16.6 kDa subunit) participates in a complex network of protein-protein interactions both within Complex I and potentially with other mitochondrial components. These interactions are critical for proper assembly, stability, and function of the respiratory apparatus.

Within Complex I, At2g33220 interacts with multiple subunits as part of the structural architecture. Based on the subunit composition information from search result , At2g33220 is one of at least 30 different subunits that compose the complete NADH-ubiquinone oxidoreductase complex in Arabidopsis thaliana. The specific interactions include:

• Interactions with Adjacent Complex I Subunits:

  • At2g33220 likely interacts directly with several neighboring subunits within its assembly module

  • Potential interaction partners include:

    • NADH-ubiquinone oxidoreductase B 17.2 kDa subunit (At3g03100)

    • NADH-ubiquinone oxidoreductase B 14 kDa subunit (At3g12260)

    • NADH-ubiquinone oxidoreductase B 18 kDa subunit (At2g02050)

  • These interactions contribute to the structural integrity of this region of Complex I

• Interactions with Assembly Factors:

  • During the biogenesis of Complex I, At2g33220 likely interacts with specific assembly factors

  • These transient interactions facilitate proper incorporation into the growing complex

  • Once assembly is complete, these interactions are replaced by the stable interactions with other Complex I subunits

• Potential Interactions with Alternative Respiratory Components:

  • Research on AtNDB2 (an alternative NADH dehydrogenase) and AtAOX1A (alternative oxidase) demonstrated functional linkage between these components

  • At2g33220, as part of Complex I, may interact with or be functionally coupled to these alternative pathway components

  • The observation that AtNDB2 overexpression alone did not enhance external NADH oxidation unless AtAOX1A was concomitantly overexpressed suggests coordinated regulation between different respiratory components

• Regulatory Interactions:

  • Protein kinases or phosphatases may interact with At2g33220 to regulate its function through phosphorylation/dephosphorylation

  • Redox-sensitive proteins may interact with At2g33220 to modulate its activity based on cellular redox state

• Interactions During Stress Responses:

  • Under stress conditions, At2g33220 may form novel interactions with stress-response proteins

  • These interactions could modulate Complex I activity to accommodate altered metabolic demands during stress

Experimental approaches to study these interactions include:

• Blue Native PAGE to preserve native complexes and identify stable interaction partners

• Co-immunoprecipitation with At2g33220-specific antibodies followed by mass spectrometry

• Yeast two-hybrid or split-GFP assays to detect binary interactions

• Crosslinking mass spectrometry to capture transient or weak interactions

• Proximity labeling techniques (BioID, APEX) to identify proteins in close proximity to At2g33220 in vivo

Understanding these interactions would provide valuable insights into how At2g33220 contributes to the assembly, stability, and regulation of respiratory complexes in plant mitochondria.

What are the knowledge gaps in understanding At2g33220 function and regulation?

Despite advances in understanding plant mitochondrial function, several significant knowledge gaps remain regarding At2g33220 function and regulation:

Addressing these knowledge gaps would provide a more comprehensive understanding of At2g33220's role in plant mitochondrial function and could reveal new approaches for enhancing plant stress resilience through targeted modifications of respiratory components.

What novel experimental approaches could advance our understanding of At2g33220 function?

Several innovative experimental approaches could significantly advance our understanding of At2g33220 function beyond conventional methods:

• Cryo-Electron Microscopy of Plant Complex I:

  • High-resolution structural analysis of plant-specific Complex I with identified subunit positions

  • Implementation of subtomogram averaging to visualize At2g33220 in its native conformation

  • Comparison of structures under different physiological conditions to detect conformation changes

  • This approach would provide unprecedented insights into At2g33220's structural contributions to Complex I

• Single-Molecule Tracking in Living Plant Cells:

  • Development of minimally disruptive fluorescent tags for At2g33220

  • Real-time visualization of protein dynamics within mitochondria

  • FRET-based approaches to monitor interactions with other respiratory components

  • This would reveal dynamic aspects of At2g33220 function impossible to observe with static techniques

• Organelle-Specific Proximity Labeling:

  • Fusion of At2g33220 with BioID, APEX, or TurboID for in vivo proximity labeling

  • Identification of the complete interactome under different conditions

  • Temporal analysis of interaction changes during stress responses

  • This would map the protein's interaction network comprehensively

• CRISPR-Mediated Base Editing:

  • Precise introduction of point mutations without double-strand breaks

  • Creation of allelic series to identify critical residues for function

  • Simultaneous editing of multiple Complex I subunits to study cooperative effects

  • This approach would enable functional dissection of specific protein domains

• In Organello Translation Systems:

  • Development of isolated mitochondria systems capable of protein import and Complex I assembly

  • Real-time monitoring of At2g33220 incorporation into Complex I

  • Identification of assembly intermediates and chaperones

  • This would reveal the biogenesis pathway of Complex I involving At2g33220

• Multi-Omics Integration with Machine Learning:

  • Simultaneous analysis of transcriptome, proteome, metabolome, and phenome data

  • Application of machine learning algorithms to identify patterns and correlations

  • Prediction of At2g33220 function under untested conditions

  • This would provide a systems-level understanding of At2g33220's role

• Optogenetic Control of At2g33220 Function:

  • Engineering light-sensitive domains into At2g33220

  • Precise temporal control of protein activity in specific tissues

  • Real-time observation of physiological consequences of activation/inactivation

  • This would allow direct testing of cause-effect relationships in vivo

• Nanoscale Respirometry:

  • Development of microfluidic devices for single-mitochondrion respiration measurements

  • Comparison of wild-type and At2g33220 mutant mitochondria at the individual organelle level

  • This would reveal heterogeneity in mitochondrial responses that are masked in bulk measurements

• Synthetic Biology Approaches:

  • Reconstruction of minimal respiratory complexes with defined subunit composition

  • Systematic addition/removal of subunits to determine essential versus accessory functions

  • This would define the minimal structural requirements for Complex I function

These innovative approaches would complement traditional biochemical and genetic methods, providing new dimensions of understanding that are currently inaccessible with conventional techniques. The combination of structural, dynamic, systems-level, and synthetic approaches would create a comprehensive picture of At2g33220 function within the complex landscape of plant mitochondrial respiration.

How might understanding At2g33220 contribute to improving plant stress resilience in changing environments?

Understanding At2g33220 could substantially contribute to improving plant stress resilience in changing environments by targeting a fundamental component of mitochondrial respiration. This knowledge could be translated into practical applications through several pathways:

The search results support the feasibility of these approaches, as studies have already demonstrated that modifications to respiratory components can significantly affect stress tolerance. For example, plants lacking either AtAOX1A or AtNDB2 showed increased sensitivity to combined drought and elevated light treatments, whereas plants overexpressing these components showed increased tolerance . Similar principles could be applied to At2g33220 engineering, potentially with even more significant effects given its central role in the main respiratory pathway rather than alternative routes.