Recombinant Lampetra tridentata Cytochrome c (cyc)

What is Lampetra tridentata cytochrome c and why is it significant for research?

Cytochrome c is a highly conserved heme protein that functions primarily in electron transport within the mitochondrial respiratory chain. Lampetra tridentata (Pacific lamprey) cytochrome c is of particular interest to researchers because lampreys represent one of the most ancient vertebrate lineages, offering unique evolutionary insights into the structure-function relationships of this essential protein. The recombinant expression of this protein allows for detailed characterization of its structural features, redox properties, and functional aspects without requiring extraction from the native organism. Like other cytochrome c proteins, it likely adopts a globular α-helical fold with His/Met coordination of the heme iron, making it valuable for comparative studies across species .

How does Lampetra tridentata cytochrome c compare structurally to other species' cytochrome c?

Lampetra tridentata cytochrome c would be expected to share the core structural elements common to all c-type cytochromes while potentially exhibiting unique evolutionary adaptations. Based on studies of other cytochrome c proteins, we can predict it contains:

• A globular α-helical fold surrounding a central heme group

• Covalent attachment of the heme via two thioether linkages to cysteine residues

• His/Met axial ligation to the heme iron

• Well-conserved amino acid residues around the heme binding site

Tertiary structural predictions could be performed by threading the primary amino acid sequence onto known crystal structures using tools such as the Phyre2 recognition engine, similar to methods used for analyzing C. elegans cytochrome c variants . For precise structural comparisons, researchers would need to perform detailed spectroscopic analyses and potentially X-ray crystallography or NMR studies.

What are the typical biophysical parameters of recombinant cytochrome c proteins?

Recombinant cytochrome c proteins typically exhibit the following biophysical parameters:

These parameters may vary for Lampetra tridentata cytochrome c based on its specific amino acid composition and evolutionary adaptations. Characterization would involve spectroscopic tools to determine structural features and stability, similar to approaches used for C. elegans cytochrome c variants .

What expression systems are most effective for producing recombinant Lampetra tridentata cytochrome c?

Based on successful cytochrome c expression systems documented in the literature, several options are available for recombinant Lampetra tridentata cytochrome c production:

• E. coli Expression with Cytochrome c Maturation Proteins: The most widely used approach involves co-expression of the cytochrome c gene with the E. coli cytochrome c maturation (Ccm) proteins (CcmABCDEFGH, System I) that assist with proper heme attachment. This can be achieved using a dual-plasmid system: one containing the target cytochrome c gene and another (such as pEC86) containing the ccm genes .

• pBTR Plasmid System: For proteins that may not express well with the dual-plasmid approach, the pBTR plasmid system (which has been successful for horse heart cytochrome c expression) could be employed. This system contains both the cytochrome c gene and the necessary maturation factors in a single plasmid .

• Yeast Expression Systems: For proteins requiring eukaryotic post-translational processing, yeast expression systems can be effective, as demonstrated with Katsuwonus pelamis cytochrome c .

• Mammalian Cell Expression: For applications requiring mammalian-specific modifications, HEK293 cells have been successfully used to express human cytochrome c .

The choice of expression system depends on research needs, with E. coli systems generally providing higher yields but yeast or mammalian systems potentially offering more native-like post-translational modifications.

How can I optimize the yield of recombinant Lampetra tridentata cytochrome c in E. coli?

To optimize yield of recombinant Lampetra tridentata cytochrome c in E. coli, consider the following evidence-based approaches:

• Media Selection: Terrific Broth (TB) media typically yields higher expression levels than standard LB media for cytochrome c proteins .

• Temperature Optimization: While initial growth at 37°C is common, reducing temperature to 25-30°C after induction can enhance proper folding and heme incorporation.

• Strategic Codon Optimization: Adapting the Lampetra tridentata cytochrome c gene sequence to E. coli codon usage preferences while preserving critical functional motifs.

• Periplasmic Targeting: Using a periplasmic targeting sequence (such as pelB) can improve heme incorporation as demonstrated with C. elegans cytochrome c .

• Culture Screening Protocol:

  • Transform plasmid(s) into appropriate E. coli strains (BL21(DE3) or derivatives)

  • Screen multiple colonies for expression (look for red-colored cell pellets indicating successful heme incorporation)

  • Perform small-scale expression tests before scaling up

  • Monitor growth at OD600 and harvest cells when cytochrome c production peaks

• Induction Optimization: Titrate IPTG concentration and induction timing to identify optimal conditions.

A systematic approach testing these variables would be necessary to determine optimal conditions specifically for Lampetra tridentata cytochrome c.

What purification strategies yield the highest purity recombinant Lampetra tridentata cytochrome c?

Based on established cytochrome c purification protocols, a multi-step purification strategy would be recommended:

• Initial Extraction:

  • Cell lysis via sonication or French press in a 50 mM Tris-HCl buffer (pH 7.5)

  • Removal of cell debris by centrifugation (20,000 × g, 30 min)

• Affinity Chromatography:

  • If the recombinant protein contains a His-tag, immobilized metal affinity chromatography (IMAC) using Ni-NTA resin is effective

  • For non-tagged protein, CM-Sepharose cation exchange chromatography can be used (cytochrome c binds due to its positive charge)

• Ion Exchange Chromatography:

  • Further purification using a high-resolution ion exchange column (e.g., MonoS) with a salt gradient elution

• Size Exclusion Chromatography:

  • Final polishing step using a Superdex 75 or similar column to separate by molecular size and remove aggregates

• Quality Assessment:

  • SDS-PAGE analysis to confirm >90-95% purity

  • Spectroscopic analysis of the A410/A280 ratio to assess heme incorporation

  • Mass spectrometry to confirm molecular weight and integrity

This multi-step approach typically yields cytochrome c preparations with >95% purity suitable for detailed biochemical and biophysical characterization.

What spectroscopic methods are most informative for characterizing recombinant Lampetra tridentata cytochrome c?

Multiple complementary spectroscopic methods are recommended for comprehensive characterization:

• UV-Visible Absorption Spectroscopy:

  • Primary method for assessing heme environment and oxidation state

  • Oxidized cytochrome c: strong Soret band (~410 nm) and weaker bands at ~530 nm and ~695 nm

  • Reduced cytochrome c: Soret band shift to ~415 nm and sharp α and β bands at ~550 nm and ~520 nm

  • The 695 nm band specifically indicates Met-Fe ligation characteristic of native cytochrome c

• Circular Dichroism (CD) Spectroscopy:

  • Far-UV CD (190-250 nm): Quantifies secondary structure content (α-helices, β-sheets)

  • Near-UV CD (250-350 nm): Provides information on tertiary structure

  • Visible CD: Reports on the heme environment configuration

• Fluorescence Spectroscopy:

  • Intrinsic tryptophan fluorescence to monitor tertiary structure changes

  • Time-resolved fluorescence resonance energy transfer (TR-FRET) using strategically placed fluorescent labels to assess conformational dynamics

• Nuclear Magnetic Resonance (NMR) Spectroscopy:

  • 1D and 2D NMR for detailed structural analysis of the protein in solution

  • Particularly valuable for mapping the heme pocket environment

• Resonance Raman Spectroscopy:

  • Provides specific information about the heme iron coordination and spin state

  • Can distinguish between different axial ligand configurations

Each method provides complementary information, and combining these approaches allows for comprehensive structural characterization of recombinant Lampetra tridentata cytochrome c.

How can I accurately determine the redox potential of recombinant Lampetra tridentata cytochrome c?

Accurate determination of the redox potential is crucial for understanding the functional properties of cytochrome c. The following methodological approach is recommended:

• Spectroelectrochemical Titration:

  • Prepare protein sample (typically 5-10 μM) in buffer containing redox mediators (e.g., methylene blue, phenazine methosulfate)

  • Use a thin-layer spectroelectrochemical cell with a three-electrode system

  • Apply controlled potentials while monitoring absorbance changes at characteristic wavelengths (550 nm for reduced form)

  • Calculate the redox potential using the Nernst equation from the absorbance vs. potential plot

• Cyclic Voltammetry:

  • Immobilize protein on a modified electrode surface (e.g., gold electrode with self-assembled monolayers)

  • Perform cyclic voltammetry at different scan rates

  • Analyze the peak potentials to determine formal reduction potential

  • Reference against standard hydrogen electrode (SHE)

• Differential Pulse Voltammetry:

  • Higher sensitivity alternative to cyclic voltammetry

  • Allows detection of redox processes with smaller current signals

• Equilibrium Redox Titration:

  • Titrate protein with reducing agents (e.g., sodium dithionite) or oxidizing agents (e.g., potassium ferricyanide)

  • Monitor spectral changes and use reference dyes with known potentials

The redox potential should be measured under standardized conditions (pH 7.0, 25°C) and reported relative to the standard hydrogen electrode (SHE) to allow comparison with other cytochrome c species .

What methods can reveal the interaction between recombinant Lampetra tridentata cytochrome c and cardiolipin-containing membranes?

Based on established protocols for studying cytochrome c-cardiolipin interactions, the following methods would be most informative:

• Liposome Binding Assays:

  • Prepare liposomes of defined composition containing cardiolipin (CL)

  • Incubate cytochrome c with liposomes and separate bound/unbound fractions

  • Quantify binding using spectroscopic methods or fluorescence

  • Determine binding constants and stoichiometry

• Peroxidase Activity Measurements:

  • CL binding induces conformational changes that enhance peroxidase activity

  • Monitor peroxidase activity using substrates like Amplex Red or ABTS

  • Compare activity of free vs. CL-bound cytochrome c

  • Kinetic analysis provides insights into the functional consequences of binding

• Time-Resolved FRET (TR-FRET):

  • Label cytochrome c with appropriate fluorophores (e.g., bimane) at strategic positions

  • Measure dye-to-heme distance distributions in the presence/absence of CL-containing liposomes

  • Analyze for the presence of extended and compact conformer populations

  • This approach has revealed distinct conformational ensembles in CL-bound cytochrome c from other species

• Circular Dichroism Spectroscopy:

  • Monitor changes in secondary structure upon CL binding

  • Quantify helical content alterations that typically accompany membrane interaction

• Atomic Force Microscopy:

  • Visualize cytochrome c interaction with supported lipid bilayers containing CL

  • Observe protein clustering or membrane perturbations

These approaches would reveal whether Lampetra tridentata cytochrome c exhibits similar membrane interaction properties to those observed in other species, providing insights into the evolutionary conservation of this function.

How can mutagenesis of Lampetra tridentata cytochrome c inform structure-function relationships?

Site-directed mutagenesis provides a powerful approach to investigate structure-function relationships in Lampetra tridentata cytochrome c. Based on approaches with other cytochrome c proteins, consider the following strategy:

• Target Selection Based on Sequence Alignment:

  • Identify conserved residues across species and lamprey-specific residues

  • Focus on:

    • Heme pocket residues (particularly axial ligands His18 and Met80, using horse numbering)

    • Surface residues involved in binding to physiological partners

    • Residues unique to Lampetra compared to other vertebrates

• Strategic Mutation Types:

  • Conservative mutations to probe subtle effects (e.g., His→Asn, Met→Leu)

  • Non-conservative mutations to significantly alter properties (e.g., charged→hydrophobic)

  • Insertion of non-natural amino acids for specialized probes

• Functional Analyses:

  • Measure redox potential changes in mutants

  • Assess thermal and chemical stability using circular dichroism

  • Compare peroxidase activity in the presence/absence of cardiolipin

  • Evaluate electron transfer kinetics with physiological partners

• Structure Determination:

  • Use X-ray crystallography or NMR to determine structural consequences of mutations

  • Apply computational modeling to predict and analyze structural changes

This approach has been successfully applied to cytochrome c from various species to understand the molecular basis of redox properties, stability, and interaction with binding partners .

What can evolutionary analysis of Lampetra tridentata cytochrome c reveal about protein function?

Evolutionary analysis of Lampetra tridentata cytochrome c can provide valuable insights into protein function through several approaches:

• Phylogenetic Analysis:

  • Construct phylogenetic trees including cytochrome c sequences from Lampetra and other vertebrates and invertebrates

  • Identify lineage-specific adaptations and conserved regions

  • Calculate evolutionary rates for different protein regions

  • Map evolutionary conservation onto the protein structure

• Ancestral Sequence Reconstruction:

  • Infer ancestral cytochrome c sequences at key evolutionary nodes

  • Express and characterize reconstructed ancestral proteins

  • Compare functional properties of ancestral and extant proteins

• Positive Selection Analysis:

  • Use statistical methods (e.g., dN/dS ratios) to identify sites under positive selection

  • Correlate selected sites with functional regions and known interactions

  • Test hypotheses about adaptive evolution through experimental characterization

• Structural Comparison Across Lineages:

  • Compare tertiary structures of cytochrome c from lampreys, jawed vertebrates, and invertebrates

  • Identify structural elements that have remained conserved despite sequence divergence

  • Create a structure-based sequence alignment to reveal functionally important residues

As lampreys represent one of the most ancient vertebrate lineages, Lampetra tridentata cytochrome c provides a unique window into the early evolution of vertebrate mitochondrial proteins and their functional adaptations .

How can recombinant Lampetra tridentata cytochrome c be used to study apoptotic mechanisms?

Recombinant Lampetra tridentata cytochrome c offers a valuable tool for comparative studies of apoptotic mechanisms across evolutionary lineages, especially given the controversial role of cytochrome c in invertebrate apoptosis . The following research approaches would be particularly informative:

• Comparative Apoptotic Activity Assays:

  • Cell-free systems using cytoplasmic extracts to assess apoptosome formation

  • Compare the ability of Lampetra cytochrome c to activate caspases relative to mammalian cytochrome c

  • Quantify apoptogenic potential using reconstituted systems with purified components

• Structure-Function Analysis of Apoptotic Activity:

  • Create chimeric proteins combining domains from Lampetra and mammalian cytochrome c

  • Identify structural elements responsible for differential apoptotic activity

  • Use mutagenesis to map the specific residues critical for apoptosome activation

• Cardiolipin Interaction Studies:

  • Characterize the interaction between Lampetra cytochrome c and cardiolipin-containing membranes

  • Measure peroxidase activity enhancement upon cardiolipin binding

  • Compare conformational dynamics using TR-FRET analysis as described for C. elegans cytochrome c

  • Analyze extended and compact conformer populations in the presence of cardiolipin

• Cross-Species Compatibility Testing:

  • Test whether Lampetra cytochrome c can substitute for mammalian cytochrome c in apoptotic pathways

  • Perform rescue experiments in cytochrome c-depleted cellular systems

These approaches would help elucidate when and how cytochrome c's role in apoptosis evolved and potentially reveal novel aspects of the protein's function in early vertebrates.

What are common challenges in recombinant cytochrome c expression and how can they be addressed?

Several challenges are commonly encountered in recombinant cytochrome c expression, with corresponding solutions:

Early detection of these issues through small-scale test expressions and appropriate spectroscopic analysis can save significant time and resources in the production of high-quality recombinant cytochrome c.

How can I validate that recombinant Lampetra tridentata cytochrome c is correctly folded with proper heme incorporation?

Comprehensive validation of proper folding and heme incorporation requires multiple complementary approaches:

• Spectroscopic Validation:

  • UV-visible absorption spectroscopy:

    • Correctly folded ferric (oxidized) cytochrome c shows characteristic Soret band at ~410 nm

    • The presence of a weak absorption band at ~695 nm confirms Met-Fe coordination

    • Reduced form exhibits sharp α band at ~550 nm and β band at ~520 nm

  • A410/A280 ratio >4 indicates high heme incorporation efficiency

• Functional Validation:

  • Electron transfer activity with physiological partners

  • Expected redox potential range (+200 to +350 mV vs. SHE)

  • Peroxidase activity enhancement upon cardiolipin binding

• Structural Validation:

  • Circular dichroism to confirm α-helical content characteristic of cytochrome c

  • Thermal denaturation profile with cooperative unfolding transition

  • Native PAGE showing compact migration pattern

  • Size exclusion chromatography elution profile consistent with monomeric protein

• Chemical Validation:

  • Mass spectrometry to confirm:

    • Expected molecular mass (accounting for heme attachment)

    • Correct covalent thioether bonds between heme and protein

  • Heme stain analysis following SDS-PAGE separation

Combinations of these approaches provide comprehensive evidence for proper folding and heme incorporation, essential for reliable downstream applications and interpretations.

What experimental controls are essential when studying recombinant Lampetra tridentata cytochrome c function?

Rigorous experimental design requires appropriate controls to ensure valid and interpretable results:

• Protein Quality Controls:

  • Freshly prepared vs. stored protein comparison to assess stability

  • Multiple protein preparations to ensure reproducibility

  • Concentration-dependent measurements to identify aggregation effects

  • SDS-PAGE and spectroscopic analysis before each experiment to confirm integrity

• Functional Assay Controls:

  • Commercial horse heart or yeast cytochrome c as reference standards

  • Apo-cytochrome c (heme-free) to distinguish protein vs. heme effects

  • Heat-denatured protein as negative control

  • Buffer-only controls to account for assay artifacts

• Specificity Controls:

  • Site-directed mutants affecting key functional sites

  • Competition assays with known binding partners

  • Varied reaction conditions (pH, ionic strength) to confirm physiological relevance

• For Evolutionary Studies:

  • Include cytochrome c from multiple species representing different evolutionary distances

  • Prepare all proteins using identical expression and purification protocols

  • Normalize activity to protein concentration and heme content

• For Membrane Interaction Studies:

  • Liposomes with and without cardiolipin

  • Varied cardiolipin percentages to establish dose-dependence

  • Alternative anionic lipids to test specificity of interactions

These controls help distinguish specific biological effects from artifacts and provide necessary context for interpreting experimental results in the broader framework of cytochrome c evolution and function .

How can recombinant Lampetra tridentata cytochrome c be used to develop novel biosensors?

Recombinant Lampetra tridentata cytochrome c offers several promising avenues for biosensor development based on its inherent properties:

• Electrochemical Biosensors:

  • Direct electron transfer between cytochrome c and electrode surfaces

  • Potential applications in detecting:

    • Superoxide and reactive oxygen species

    • Cardiolipin and membrane lipid composition

    • Cyanide and other heme-binding toxins

  • Implementation through:

    • Protein immobilization on nanomaterials (carbon nanotubes, graphene)

    • Self-assembled monolayer techniques

    • Incorporation into biocompatible polymers

• Peroxidase-Based Optical Biosensors:

  • Exploit the peroxidase activity enhanced by cardiolipin binding

  • Design fluorogenic or chromogenic assays for H₂O₂ detection

  • Applications in oxidative stress monitoring in biological systems

• Conformational Change-Based Biosensors:

  • Engineer FRET-based sensors using the conformational changes that occur upon:

    • Redox state changes

    • Cardiolipin binding

    • pH variations

  • These could serve as intracellular probes for mitochondrial status

The evolutionary distinctiveness of Lampetra tridentata cytochrome c might provide unique properties (stability, substrate specificity, or redox characteristics) advantageous for specific biosensing applications compared to mammalian cytochrome c.

What are the key considerations for studying post-translational modifications of recombinant Lampetra tridentata cytochrome c?

Investigating post-translational modifications (PTMs) of recombinant Lampetra tridentata cytochrome c requires careful attention to several key factors:

• Expression System Selection:

  • E. coli systems: Suitable for studying the native unmodified protein but lack eukaryotic PTM machinery

  • Yeast systems: Provide some eukaryotic PTMs but may differ from native modifications

  • Mammalian expression systems: Offer most authentic eukaryotic PTMs but with lower yields

• PTM Identification Strategy:

  • Mass spectrometry approaches:

    • Bottom-up proteomics: Enzymatic digestion followed by LC-MS/MS

    • Top-down proteomics: Analysis of intact protein to preserve modification relationships

    • Targeted MS methods for known modifications (e.g., MRM for phosphorylation sites)

  • Site-specific antibodies for common PTMs (phosphorylation, acetylation)

  • Specialized staining methods (Pro-Q Diamond for phosphorylation)

• Functional Impact Assessment:

  • Compare modified vs. unmodified protein for:

    • Redox potential differences

    • Stability changes

    • Altered binding to physiological partners

    • Modified peroxidase activity and cardiolipin interaction

• Physiological Context:

  • Determine whether modifications occur in response to specific cellular conditions

  • Assess evolutionary conservation of modification sites across species

  • Investigate enzymes responsible for adding/removing modifications

Known cytochrome c PTMs in other species include phosphorylation, acetylation, and oxidative modifications, all of which can significantly impact function. Studying these in the context of an ancient vertebrate lineage could provide evolutionary insights into the emergence and significance of these regulatory mechanisms.

How might differential expression systems affect the properties of recombinant Lampetra tridentata cytochrome c?

Different expression systems can significantly impact the properties of recombinant cytochrome c, with important considerations for research applications:

To determine the optimal expression system for specific research goals:

• For structural or basic biochemical studies where high yield is prioritized, E. coli systems are generally preferred .

• For studies investigating native PTMs or applications requiring highly authentic protein, mammalian or yeast systems would be more appropriate despite lower yields .

• For comprehensive characterization, comparing proteins from multiple expression systems can provide valuable insights into the impact of expression context on protein properties.

The choice of expression system should be guided by the specific research questions being addressed and the properties most critical to maintain for the particular application.

What are the recommended storage conditions to maintain recombinant Lampetra tridentata cytochrome c stability?

Based on established protocols for cytochrome c storage, the following evidence-based recommendations apply:

• Short-term Storage (1-2 weeks):

  • Store at 4°C in appropriate buffer:

    • 20-50 mM sodium phosphate or Tris buffer, pH 7.0-7.5

    • 100-150 mM NaCl to maintain ionic strength

  • Add reducing agent (e.g., 1 mM DTT or 2 mM β-mercaptoethanol) to prevent Met80 oxidation

  • Filter-sterilize (0.22 μm) to prevent microbial growth

• Medium-term Storage (1-6 months):

  • Aliquot to avoid freeze-thaw cycles

  • Store at -20°C with 10-20% glycerol as cryoprotectant

  • Seal tubes under nitrogen if possible to prevent oxidation

• Long-term Storage (>6 months):

  • Lyophilize in the presence of stabilizing agents (e.g., trehalose)

  • Store at -80°C with desiccant

  • Alternatively, store concentrated solutions (>1 mg/mL) at -80°C in small aliquots

• Stability Monitoring Protocol:

  • Before each experiment, check:

    • UV-visible spectrum (A410/A280 ratio and 695 nm band)

    • Functional activity (electron transfer or peroxidase assay)

  • Document any changes in spectral properties or activity over time

Following these protocols minimizes protein degradation, oxidative damage, and denaturation, ensuring reliable and reproducible experimental results.

What standardized reporting should be included when publishing research using recombinant Lampetra tridentata cytochrome c?

When publishing research using recombinant Lampetra tridentata cytochrome c, the following standardized reporting elements should be included to ensure reproducibility and proper interpretation:

• Expression and Purification Details:

  • Complete gene sequence or database accession number

  • Expression vector and host system details

  • Inclusion of tags or fusion partners and whether they were removed

  • Detailed purification protocol with column types and buffer compositions

  • Final purity assessment method and percentage purity achieved

• Protein Characterization:

  • UV-visible spectral properties (A410/A280 ratio, presence of 695 nm band)

  • Mass spectrometry confirmation of molecular weight

  • Heme content quantification method and results

  • Circular dichroism data if structural characterization was performed

  • Redox potential with reference electrode and measurement conditions

• Experimental Conditions:

  • Buffer composition, pH, and ionic strength

  • Temperature and any other relevant environmental factors

  • Protein concentration determination method

  • Sample handling procedures

  • Time between purification and experimentation

• Controls and Standards:

  • Comparison with commercial cytochrome c standards when applicable

  • Positive and negative controls for functional assays

  • Statistical analysis methods and significance criteria

• Data Accessibility:

  • Deposition of sequence data in appropriate databases

  • Sharing of raw spectral or experimental data in repositories

  • Availability of plasmids or strains to the research community

This comprehensive reporting ensures that results can be properly evaluated, compared with other studies, and reproduced by other researchers in the field.

What are the most promising future research directions for Lampetra tridentata cytochrome c studies?

Based on current trends in cytochrome c research and the unique evolutionary position of Lampetra tridentata, several promising research directions emerge:

• Evolutionary Biochemistry:

  • Detailed comparison of Lampetra tridentata cytochrome c with counterparts from jawless, jawed vertebrates, and invertebrates

  • Ancestral sequence reconstruction and characterization to understand the trajectory of cytochrome c evolution

  • Investigation of selection pressures on different protein regions throughout vertebrate evolution

• Structure-Function Relationship Insights:

  • High-resolution structural determination (X-ray crystallography or cryo-EM)

  • Dynamics studies using NMR or hydrogen-deuterium exchange mass spectrometry

  • Computational simulations to understand the molecular basis of functional properties

• Apoptosis Regulation Studies:

  • Comparative analysis of apoptosome activation capacity

  • Investigation of cardiolipin interaction specifics and peroxidase activity

  • Exploration of lamprey-specific apoptotic pathways and cytochrome c involvement

• Applications in Biotechnology:

  • Development of stable cytochrome c variants for biosensor applications

  • Exploration of unique catalytic properties for biocatalysis

  • Investigation of potential therapeutic applications based on unique properties

• Systems Biology Approaches:

  • Integration of cytochrome c into larger networks of mitochondrial function

  • Comparative mitochondrial proteomics across evolutionary lineages

  • Investigation of regulatory mechanisms in primitive vertebrates