Recombinant Nitrosomonas europaea Putative iron-sulfur cluster insertion protein ErpA (erpA)

What is Nitrosomonas europaea and why is it significant for iron-sulfur cluster research?

Nitrosomonas europaea is a gram-negative obligate chemolithoautotroph that derives all its energy and reducing power from the oxidation of ammonia to nitrite, playing a crucial role in the biogeochemical nitrogen cycle through nitrification. Its genome consists of a single circular chromosome of 2,812,094 bp with approximately 2,460 protein-encoding genes .

The significance of N. europaea in iron-sulfur cluster research stems from its unique iron acquisition strategy. The organism has evolved sophisticated mechanisms to accumulate iron from its environment, with more than 20 genes devoted to various classes of iron receptors . This extensive iron uptake machinery suggests that iron plays critical roles in its metabolism, particularly in the formation and maintenance of iron-sulfur clusters in proteins like ErpA. The organism's ability to thrive in iron-limited environments provides valuable insights into iron-sulfur cluster biogenesis and insertion under stress conditions.

What is the function of iron-sulfur cluster insertion proteins in bacteria?

Iron-sulfur cluster insertion proteins, like ErpA, are specialized proteins that facilitate the transfer of preformed iron-sulfur clusters from scaffold proteins to target apo-proteins. These proteins function as part of a larger iron-sulfur cluster biogenesis system that typically includes:

• Scaffold proteins (like IscU or SufB) where clusters are initially assembled

• Carrier proteins (like IscA, SufA, or ErpA) that transfer clusters to target proteins

• Target apo-proteins that require iron-sulfur clusters for their function

In E. coli, a close relative to the systems studied in N. europaea, iron-sulfur clusters are built on either IscU or SufB scaffolds and then delivered to target proteins via carrier proteins like IscA and SufA . ErpA likely plays a similar role in N. europaea, serving as an A-type carrier (ATC) protein that facilitates the final transfer of iron-sulfur clusters to target proteins under specific environmental conditions.

The precise delivery pathways can vary depending on conditions, with differences arising in the [Fe-S] delivery steps between different target proteins .

How does the ErpA protein contribute to iron-sulfur cluster biogenesis in Nitrosomonas europaea?

Based on comparative analyses with homologous systems, the putative ErpA protein in N. europaea likely serves as a specialized iron-sulfur cluster carrier protein that functions in the final stages of cluster transfer to specific target proteins. While the specific targets of N. europaea ErpA have not been fully characterized, research on related systems suggests that ErpA may be particularly important under iron limitation or oxidative stress conditions.

In N. europaea, which has evolved extensive iron acquisition mechanisms including multiple iron receptors , ErpA may play a crucial role in ensuring that essential iron-sulfur proteins receive their clusters even when iron availability is limited. The protein likely coordinates with other components of the iron-sulfur cluster assembly machinery to maintain cellular functions dependent on iron-sulfur proteins under varying environmental conditions.

What are the recommended protocols for cloning and expressing recombinant N. europaea ErpA protein?

For successful cloning and expression of recombinant N. europaea ErpA protein, researchers should consider the following methodological approach:

Cloning Strategy:

• Amplify the erpA gene from N. europaea genomic DNA using high-fidelity DNA polymerase

• Design primers with appropriate restriction sites compatible with your expression vector

• For optimal expression, consider codon optimization for the host organism (typically E. coli)

• Include a purification tag (His-tag or GST-tag) at either N- or C-terminus

• Clone into an expression vector with an inducible promoter (T7 or tac)

Expression Conditions:

• Transform the construct into an E. coli expression strain capable of iron-sulfur cluster assembly (BL21(DE3) or specialized strains)

• Culture in rich media (LB or TB) supplemented with iron (50-100 μM ferric ammonium citrate)

• Grow cells at 37°C until OD600 reaches 0.6-0.8

• Reduce temperature to 16-18°C before induction

• Induce with 0.1-0.5 mM IPTG

• Continue expression for 16-20 hours at reduced temperature

Purification Considerations:

• Perform all purification steps under anaerobic conditions to preserve iron-sulfur clusters

• Include reducing agents (5-10 mM β-mercaptoethanol or 1-2 mM DTT) in all buffers

• Consider including iron and sulfide in buffers (5-10 μM each) to stabilize clusters

• Use affinity chromatography followed by size exclusion chromatography

This approach maximizes the likelihood of obtaining functional recombinant ErpA protein with intact iron-sulfur clusters for subsequent biochemical and structural analyses.

How can researchers verify the presence and integrity of iron-sulfur clusters in purified recombinant ErpA?

Verifying the presence and integrity of iron-sulfur clusters in purified recombinant ErpA requires multiple complementary analytical techniques:

Spectroscopic Methods:

• UV-visible absorption spectroscopy: Iron-sulfur proteins typically show broad absorption bands at 320-450 nm. Monitor the ratio of absorbance at 410 nm to 280 nm to assess cluster content.

• Circular dichroism (CD) spectroscopy: Iron-sulfur clusters exhibit characteristic CD spectra in the visible region (300-700 nm).

• Electron paramagnetic resonance (EPR) spectroscopy: For characterizing the redox state and environment of the iron-sulfur cluster.

Biochemical Analysis:

• Iron and sulfide quantification: Determine the Fe:S:protein ratio using colorimetric assays.

  • Iron content: Use ferene method or atomic absorption spectroscopy

  • Sulfide content: Use methylene blue method

  • Protein content: Bradford or BCA assay

Functional Assays:

• Cluster transfer assays: Monitor the ability of purified ErpA to transfer clusters to known target proteins.

• Electron transfer assays: If applicable, measure electron transfer capabilities using redox-active dyes.

Stability Assessment:

• Monitor cluster integrity under various conditions (temperature, pH, oxidants)

• Track cluster loss during storage using UV-visible spectroscopy

A typical iron-sulfur cluster content for properly folded [2Fe-2S] or [4Fe-4S] A-type carrier proteins should yield approximately 1 cluster per protein monomer, with a characteristic brownish color of the protein solution indicating the presence of intact clusters.

What expression systems are most effective for producing functional recombinant iron-sulfur proteins like ErpA?

The choice of expression system significantly impacts the successful production of functional iron-sulfur proteins. Based on research with similar proteins, the following systems have proven effective:

E. coli-Based Systems:

• E. coli BL21(DE3) with co-expression of iron-sulfur cluster assembly machinery

  • Include plasmids encoding the isc or suf operon components

  • Supplement media with iron and cysteine sources

• E. coli strains with enhanced cytoplasmic disulfide bond formation (e.g., Origami strains)

  • Helps maintain proper protein folding

• Specialized E. coli strains like CyaY-knockout strains

  • May enhance iron-sulfur cluster formation in heterologous proteins

Alternative Prokaryotic Systems:

• Bacterial hosts with high iron tolerance and natural iron uptake systems

• Anaerobic expression systems to prevent cluster oxidation

Expression Parameters to Optimize:

• Temperature: Typically lower temperatures (16-20°C) improve folding and cluster incorporation

• Induction: Use low inducer concentrations for slower expression

• Media composition: Supplement with iron (50-100 μM) and sulfur sources

• Growth conditions: Consider microaerobic or anaerobic cultivation

Comparative Expression Efficiency Table:

The most effective strategy typically involves optimized E. coli expression with co-expression of iron-sulfur cluster assembly machinery at reduced temperatures, combined with careful anaerobic purification procedures.

How does the structure and function of ErpA in N. europaea compare with homologous proteins in other bacteria?

The comparison of ErpA across different bacterial species reveals important structural and functional conservation as well as adaptations specific to each organism's ecological niche:

Structural Conservation:

N. europaea-Specific Adaptations:

• N. europaea ErpA likely contains adaptations optimized for functioning in a chemolithoautotrophic lifestyle

• Its sequence may reflect specialization for functioning under the fluctuating iron conditions experienced in nitrifying environments

• The protein likely contains surface features that enable specific interactions with partner proteins involved in the organism's unique metabolic pathways

Functional Comparisons:
While E. coli and other model organisms often contain multiple A-type carrier proteins (IscA, SufA, ErpA) with partially overlapping functions, the specialization of these proteins in N. europaea may differ based on its unique iron acquisition strategy involving multiple iron receptors and limited siderophore production capabilities .

In E. coli, ErpA has been shown to be specifically involved in the maturation of certain respiratory complexes and is essential under aerobic conditions. The N. europaea homolog may play similar roles, but potentially with adaptations related to the organism's ammonia oxidation-based energy metabolism.

What role might ErpA play in N. europaea's adaptation to iron limitation conditions?

ErpA may play crucial roles in the adaptation to iron limitation through several mechanisms:

• Priority Distribution: Under iron limitation, ErpA might prioritize cluster delivery to essential iron-sulfur proteins while non-essential pathways are downregulated

• Enhanced Cluster Stability: The protein may have evolved to maintain cluster integrity under low iron conditions for longer periods than homologs from iron-replete environments

• Alternative Cluster Types: ErpA might facilitate the incorporation of alternative forms of iron-sulfur clusters that require less iron (e.g., [2Fe-2S] instead of [4Fe-4S])

• Integration with Iron Sensing: ErpA function may be coordinated with iron sensing mechanisms to rapidly respond to changes in iron availability

• Biofilm Context: Given that N. europaea forms significantly more biovolume when co-cultured with P. aeruginosa than in single-species biofilms , ErpA might function differently in biofilm contexts where iron availability and utilization patterns differ from planktonic growth

These adaptations would be consistent with N. europaea's environmental niche and metabolic requirements, where iron-sulfur proteins are likely essential for energy generation through ammonia oxidation, a process that must be maintained even under suboptimal iron conditions.

How can researchers investigate the in vivo interaction network of ErpA in N. europaea?

Investigating the in vivo interaction network of ErpA in N. europaea requires specialized approaches due to the protein's role in iron-sulfur cluster transfer and the unique physiology of this chemolithoautotrophic bacterium. The following methodological approaches can be employed:

Genetic Approaches:

• Construct tagged versions of ErpA (e.g., with a small epitope tag that minimally disrupts function)

• Develop a controllable expression system for N. europaea, which may require adaptation of existing tools for this non-model organism

• Create conditional mutants using techniques like CRISPR interference (CRISPRi) to deplete ErpA and identify affected pathways

Protein-Protein Interaction Methods:

• In vivo crosslinking followed by mass spectrometry (XL-MS)

  • Use cell-permeable crosslinkers that can capture transient interactions

  • Apply varying crosslinker concentrations and reaction times

  • Identify crosslinked peptides by LC-MS/MS analysis

• Proximity-based labeling approaches

  • Fuse ErpA to enzymes like BioID or APEX2

  • Express in N. europaea under different growth conditions

  • Identify labeled proteins by streptavidin pulldown and MS

• Co-immunoprecipitation from N. europaea cell lysates

  • Prepare lysates under anaerobic conditions to preserve interactions

  • Use specific antibodies against ErpA or epitope tags

  • Analyze by LC-MS/MS to identify co-precipitated proteins

Systems Biology Approaches:

• Transcriptome analysis comparing wild-type and ErpA-depleted strains

• Proteome profiling to identify proteins whose abundance changes upon ErpA depletion

• Metalloproteomics to characterize changes in the iron-sulfur proteome

Physiological Validation:

• Growth phenotyping under different iron concentrations

• Biofilm formation assays in pure and mixed cultures

• Activity measurements of known iron-sulfur enzymes

These approaches should be performed under varying conditions including:

• Iron replete vs. iron limited conditions

• Pure culture vs. co-culture with organisms like P. aeruginosa

• Planktonic growth vs. biofilm conditions

The resulting data can be integrated to construct a comprehensive interaction network that reveals the functional role of ErpA in iron-sulfur cluster distribution within the cell under different physiological conditions.

What are common challenges in working with recombinant ErpA and how can they be addressed?

Researchers working with recombinant ErpA from N. europaea often encounter several technical challenges. Here are the most common issues and recommended solutions:

Challenge 1: Low Expression Yield

• Cause: Toxicity due to iron-sulfur protein overexpression or codon usage differences

• Solutions:

  • Reduce expression temperature to 16-18°C

  • Use lower inducer concentrations (0.05-0.1 mM IPTG)

  • Consider codon-optimized synthetic genes

  • Use tightly controlled expression systems (e.g., pBAD)

  • Transform into specialized E. coli strains designed for toxic protein expression

Challenge 2: Poor Iron-Sulfur Cluster Incorporation

• Cause: Insufficient iron availability or oxidative damage during expression/purification

• Solutions:

  • Supplement expression media with 50-100 μM ferric ammonium citrate

  • Add L-cysteine (0.5-1 mM) as sulfur source

  • Co-express iron-sulfur assembly machinery (isc or suf operon)

  • Perform all purification steps under strictly anaerobic conditions

  • Include reducing agents (5 mM β-mercaptoethanol or 1-2 mM DTT) in all buffers

Challenge 3: Protein Instability and Aggregation

• Cause: Improper folding or cluster loss leading to aggregation

• Solutions:

  • Include stabilizing agents in buffers (10% glycerol, 150-300 mM NaCl)

  • Add low concentrations of detergents (0.05% Triton X-100)

  • Optimize pH conditions (typically pH 7.5-8.0 works well)

  • Include iron and sulfide in storage buffers (5-10 μM each)

  • Store protein under anaerobic conditions at 4°C for short term or flash-freeze in liquid nitrogen for long-term storage

Challenge 4: Inconsistent Activity in Functional Assays

• Cause: Heterogeneous protein preparation with variable cluster content

• Solutions:

  • Use additional purification steps like ion exchange chromatography

  • Employ analytical techniques to confirm cluster content before assays

  • Standardize protein:cluster ratios when comparing different preparations

  • Include positive controls with well-characterized iron-sulfur proteins

Troubleshooting Decision Tree:

• Is the protein expressing at all?

  • No → Check construct design, codon usage, toxicity issues

  • Yes → Proceed to next question

• Is the protein soluble but lacking color?

  • Yes → Iron-sulfur cluster assembly issue: address iron/sulfur availability, oxidation issues

  • No → Protein folding issue: optimize expression conditions, consider fusion partners

• Is the protein colored but unstable?

  • Yes → Cluster stability issue: optimize buffer conditions, anaerobic handling

  • No → Proceed to functional characterization

• Does the protein show inconsistent activity?

  • Yes → Heterogeneity issue: improve purification, standardize cluster content

  • No → Protein is suitable for detailed biochemical studies

This systematic approach helps identify and address specific challenges at each stage of working with recombinant ErpA.

How can researchers distinguish between ErpA-mediated phenotypes and those caused by other iron-sulfur cluster assembly proteins?

Distinguishing between phenotypes specifically associated with ErpA dysfunction versus those arising from defects in other iron-sulfur cluster assembly proteins requires carefully designed experimental approaches:

Genetic Approaches:

• Selective gene targeting: Use precise genetic tools like CRISPR/Cas9 to create specific mutations in erpA while leaving other iron-sulfur assembly genes intact

• Complementation studies: Express wild-type erpA or mutant variants in erpA-deficient strains to confirm phenotype causality

• Conditional expression systems: Develop titratable promoters for N. europaea to create varying levels of ErpA depletion

Biochemical Validation:

• Substrate specificity analysis: Identify target proteins that specifically receive clusters from ErpA versus other A-type carriers

• In vitro reconstitution experiments: Reconstitute partial assembly pathways with defined components to test specific transfer steps

• Protein-protein interaction mapping: Compare interaction networks of different A-type carriers to identify unique ErpA partners

Physiological Profiling:

• Growth condition specificity: Test phenotypes under conditions that preferentially affect different iron-sulfur assembly pathways:

  • Aerobic vs. anaerobic (affects oxidative stress on clusters)

  • Iron-replete vs. iron-limited (differentiates between redundant pathways)

  • Different carbon/nitrogen sources (affects metabolic demand for clusters)

• Target enzyme activity profiling: Measure the activity of specific iron-sulfur enzymes known to depend on different assembly factors:

  • Respiratory chain complexes (often ErpA-dependent)

  • Housekeeping enzymes (may use multiple pathways)

  • Stress-response proteins (often pathway-specific)

Distinctive Phenotypic Profiles:

By systematically applying these approaches, researchers can build a comprehensive profile of ErpA-specific functions distinct from the roles of other iron-sulfur cluster assembly proteins in N. europaea.

What advanced analytical techniques can be used to study the kinetics of iron-sulfur cluster transfer by ErpA?

Studying the kinetics of iron-sulfur cluster transfer by ErpA requires sophisticated analytical techniques that can track cluster movement between proteins in real-time. The following methodologies are particularly valuable for this purpose:

Time-Resolved Spectroscopic Methods:

• UV-visible stopped-flow spectroscopy

  • Allows monitoring of cluster transfer reactions with millisecond time resolution

  • Can detect changes in the electronic absorption spectra as clusters move between proteins

  • Experimental setup: Mix purified cluster-loaded ErpA with apo-target proteins in a stopped-flow apparatus

  • Data analysis: Fit absorbance changes at characteristic wavelengths to kinetic models

• Circular dichroism (CD) stopped-flow

  • Monitors changes in the CD spectra that occur during cluster transfer

  • Provides information about cluster environment changes during transfer

  • Particularly useful for [2Fe-2S] clusters which have distinctive CD signatures

• Fluorescence resonance energy transfer (FRET)

  • Requires labeling of donor and acceptor proteins with fluorescent probes

  • Allows real-time monitoring of protein-protein interactions during cluster transfer

  • Can provide distance information between proteins during the transfer process

Mass Spectrometry-Based Approaches:

• Hydrogen-deuterium exchange mass spectrometry (HDX-MS)

  • Maps protein conformational changes during cluster transfer

  • Identifies regions undergoing structural rearrangements during the transfer process

  • Time-resolved measurements can track the progression of structural changes

• Native mass spectrometry

  • Directly observes the mass changes associated with cluster transfer

  • Can distinguish different metalloforms of proteins

  • Allows monitoring of reaction intermediates

Electrochemical Methods:

• Protein film voltammetry

  • Measures the redox properties of iron-sulfur clusters within proteins

  • Can monitor changes in redox potential as clusters move between proteins

  • Provides information about the energetics of cluster transfer

Kinetic Analysis Considerations:

For proper kinetic analysis, researchers should:

• Determine rate constants under varying conditions:

  • Different protein concentrations to distinguish between first and second-order processes

  • Various temperatures to determine activation parameters

  • Different pH values to identify ionizable groups involved in the transfer

• Consider competing reactions:

  • Cluster degradation

  • Non-specific transfer

  • Protein aggregation

• Develop mathematical models that account for:

  • Multiple transfer steps

  • Conformational changes

  • Protein-protein interaction dynamics

Experimental Workflow Table:

By combining these techniques, researchers can develop comprehensive models of the kinetic mechanism of iron-sulfur cluster transfer mediated by ErpA in N. europaea, providing insights into how this process is regulated in response to environmental changes such as iron availability.

How might ErpA function integrate with the unique metabolism of N. europaea as an ammonia oxidizer?

N. europaea's identity as an obligate chemolithoautotroph that derives all its energy from ammonia oxidation creates a unique metabolic context for ErpA function. Future research should explore these connections through several promising avenues:

Ammonia Oxidation Pathway Integration:

• The ammonia monooxygenase (AMO) complex, central to N. europaea metabolism, requires electron transfer components that may depend on iron-sulfur clusters

• The hydroxylamine oxidoreductase (HAO) pathway includes multiple electron transport proteins that potentially require ErpA-mediated cluster insertion

• Researchers should investigate whether ErpA preferentially supplies clusters to these ammonia oxidation enzymes under iron-limited conditions

Energy Conservation Mechanisms:

• N. europaea's unique energy metabolism may require specialized iron-sulfur proteins for electron transport and ATP generation

• The interaction between ErpA and these energy-conservation systems could reveal adaptations specific to chemolithoautotrophic metabolism

• Future studies should examine how ErpA function correlates with cellular energy status and ammonia oxidation rates

Integration with Carbon Fixation:

• As an autotroph, N. europaea must fix CO2, likely using iron-sulfur enzymes in the Calvin cycle

• Research should explore whether ErpA plays a role in maintaining carbon fixation capacity under iron limitation

• The coordination between nitrogen and carbon metabolism may involve iron-sulfur proteins dependent on ErpA

Stress Response Coordination:

• Ammonia oxidation generates reactive nitrogen species that can damage iron-sulfur clusters

• ErpA may have evolved specialized features to protect or repair clusters under these conditions

• The relationship between ErpA function and nitrosative stress response merits detailed investigation

Adaptation to Environmental Niches:

• N. europaea's genome reveals extensive iron acquisition mechanisms , suggesting iron plays a critical role in its ecology

• ErpA may help the organism adapt to environments with fluctuating iron availability

• Studies comparing ErpA function in N. europaea to homologs in other ammonia oxidizers could reveal niche-specific adaptations

This research direction would significantly advance our understanding of how iron-sulfur cluster assembly systems have been adapted to support specialized metabolic lifestyles in different bacterial lineages.

What potential biotechnological applications might emerge from better understanding N. europaea ErpA function?

Enhanced understanding of N. europaea ErpA function could enable several innovative biotechnological applications across environmental, industrial, and medical domains:

Wastewater Treatment Optimization:

• N. europaea plays crucial roles in nitrification during wastewater treatment

• Understanding how ErpA facilitates ammonia oxidation under varying conditions could lead to improved process control

• Engineering ErpA function might enhance N. europaea performance in treatment systems, particularly under iron-limited conditions

• The observed enhancement of N. europaea biofilm formation in co-culture with P. aeruginosa suggests that manipulating ErpA function could improve biomass retention in bioreactors

Bioremediation Applications:

• Iron-sulfur enzymes are involved in the degradation of various environmental pollutants

• Engineered ErpA variants could potentially enhance the activity of these enzymes under challenging field conditions

• Applications could include enhanced degradation of recalcitrant compounds in contaminated soils and waters

Biosensor Development:

• ErpA's iron-sensing capabilities could be exploited to develop whole-cell biosensors for:

  • Environmental iron monitoring

  • Detection of bioavailable iron in soils

  • Monitoring of iron in wastewater treatment processes

• Fusion of reporter genes to ErpA-dependent promoters could create sensitive detection systems

Protein Engineering Applications:

• Understanding the structural basis of ErpA function could inform the design of:

  • Enhanced iron-sulfur proteins for biocatalysis

  • More stable iron-sulfur enzymes for industrial applications

  • Artificial iron-sulfur proteins with novel functions

• The molecular mechanisms of cluster transfer could inspire biomimetic catalysts for energy applications

Biofilm Control Strategies:

• Given that N. europaea forms significantly more biovolume when co-cultured with P. aeruginosa , manipulating ErpA function could:

  • Enhance beneficial biofilms in engineered systems

  • Inhibit harmful biofilms in medical or industrial settings

• The development of compounds targeting iron-sulfur cluster assembly could provide novel biofilm control agents

Comparative Performance Table of Potential Applications:

These applications represent valuable opportunities to translate fundamental research on N. europaea ErpA into practical technologies addressing important environmental and industrial challenges.

How might climate change and increasing environmental stressors affect the function of iron-sulfur cluster assembly proteins like ErpA in N. europaea populations?

Climate change introduces multiple stressors that could significantly impact iron-sulfur cluster assembly processes in environmental N. europaea populations. This represents an important frontier for research with implications for ecosystem functioning and biogeochemical cycles.

Temperature Effects:

• Rising global temperatures may affect:

  • The stability of iron-sulfur clusters in ErpA and target proteins

  • The kinetics of cluster assembly and transfer reactions

  • The folding efficiency of iron-sulfur proteins

• Research should examine how temperature fluctuations influence ErpA function in N. europaea and whether thermal adaptation mechanisms exist

Changes in Iron Bioavailability:

• Climate-driven changes in:

  • Soil pH (through altered precipitation patterns)

  • Redox conditions (through flooding or drought)

  • Organic matter content (through changes in vegetation)
    Could all affect iron solubility and availability

• ErpA function might need to adapt to these changing conditions, potentially through altered expression or modified interaction networks

Interactions with Multiple Stressors:

• Climate change introduces concurrent stressors including:

  • Increased UV radiation

  • Altered nutrient availability

  • Novel pollutant exposures

  • Changes in microbial community composition

• The combined effects of these stressors on iron-sulfur cluster metabolism require investigation, particularly since N. europaea has limited metabolic flexibility as an obligate chemolithoautotroph

Ecological Community Interactions:

• Climate change may alter the relationships between N. europaea and other microorganisms:

  • The enhancement of N. europaea biofilm formation by P. aeruginosa might be affected by changing environmental conditions

  • Competition for iron with other microorganisms may intensify

  • Changes in predation pressure could affect population dynamics

• How these community changes affect ErpA function and expression warrants study

Adaptive Responses:

• Research should investigate whether N. europaea can adapt to changing conditions through:

  • Altered regulation of erpA expression

  • Modified iron acquisition strategies

  • Physiological adaptations in iron-sulfur protein usage

• The genetic diversity of erpA genes in environmental populations could provide insights into adaptation potential

Research Priorities Matrix:

This research direction connects molecular-level understanding of iron-sulfur proteins with ecosystem-level responses to global change, providing insights that span from protein biochemistry to environmental microbiology and biogeochemistry.