Recombinant Dog Cytochrome c oxidase subunit 2 (MT-CO2)
Description
Cytochrome c oxidase subunit 2 (MT-CO2) Overview
Cytochrome c oxidase (COX), also known as Complex IV, is a large transmembrane protein complex found in the mitochondria of eukaryotic cells and in the plasma membranes of bacteria . It is the terminal enzyme in the electron transport chain, which is essential for cellular respiration. MT-CO2 is one of the subunits of this enzyme complex.
Function and Significance
COX plays a vital role in energy production by catalyzing the transfer of electrons from cytochrome c to molecular oxygen . This process is coupled with the pumping of protons across the inner mitochondrial membrane, creating an electrochemical gradient that drives the synthesis of ATP, the main energy currency of the cell.
Recombinant Production
Recombinant MT-CO2 is produced using genetic engineering techniques, where the gene encoding the protein is inserted into a host organism (e.g., E. coli) to produce large quantities of the protein in vitro . The recombinant protein can then be purified for use in research applications.
Applications in Research
Recombinant Dog Cytochrome c oxidase subunit 2 (MT-CO2) can be used in various research applications, including:
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Structural studies: Determining the three-dimensional structure of the protein to understand its function and interactions with other molecules.
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Enzyme kinetics: Studying the catalytic activity of the enzyme and its response to different substrates and inhibitors.
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Antibody development: Generating antibodies that specifically recognize the dog MT-CO2 protein, which can be used for immunohistochemistry, Western blotting, and other applications .
Purchasing Information
| Code | CSB-CF015073DO |
|---|---|
| MSDS | Available upon request |
| Size | Pls inquire |
| Source | in vitro E.coli expression system |
| Other Information | KEGG: cfa:804479STRING: 9615.ENSCAFP00000030312 |
Product Specs
Note: While we prioritize shipping the format currently in stock, please specify any format requirements in your order notes for customized preparation.
Note: All proteins are shipped with standard blue ice packs unless dry ice shipping is requested in advance. Additional fees apply for dry ice shipping.
If a specific tag type is required, please inform us, and we will prioritize its development.
Target Background
- High expression in mast cell tumors is associated with increased proliferation, angiogenesis, and decreased survival. (PMID: 28467670)
- COX-2 and prostaglandin E synthase gene expression may regulate uterine cervix patency in bitches with pyometra. (PMID: 26596635)
- COX-2(+)/EGFR(+) status may enable tumor cells to evade cytotoxic immune responses. (PMID: 25964576)
- COX-2 expression is significantly higher in canine osteosarcoma cancer stem cell spheres compared to adherent cells. (PMID: 24416158)
- COX-2 expression in canine anal sac adenocarcinoma suggests potential therapeutic utility for COX-2 inhibitors. (PMID: 23778258)
- In the studied gliomas, COX-2 expression was absent, with no c-kit immunoreactivity observed, although high-grade tumors exhibited intramural vascular expression. (PMID: 22235799)
- High COX-2 expression in osteosarcoma supports the use of COX-2 inhibitors to enhance chemotherapy response. (PMID: 22633646)
- Renal COX-2 is induced in canine and feline chronic kidney disease (CKD), particularly in relation to glomerular changes. No significant correlation was found for COX-1. (PMID: 22244709)
- Mutations in the COX-2 gene may contribute to renal dysplasia in dogs. (PMID: 21346820)
- This study correlated neoplastic cyclooxygenase-2 expression with histological type, differentiation grade, proliferative activity, estrogen receptor status, and Hsp70 and p53 protein expression. (PMID: 20731190)
- This study investigated the prognostic value of COX-2 in canine malignant mammary tumors. (PMID: 19939424)
- COX-2 inhibition improves transplanted cardiac function after long-term preservation. (PMID: 16713604)
- TonEBP/NFAT5 mediates hypertonic-induced COX-2 expression, highlighting osmoprotection's dependence on COX-2 protein levels. (PMID: 19146830)
- This study reports COX-2 expression in canine normal and neoplastic mammary glands. (PMID: 19203768)
KEGG: cfa:804479
STRING: 9615.ENSCAFP00000030312
Q&A
Basic Research Questions
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What is MT-CO2 and what is its functional role in canine cellular respiration?
MT-CO2 (mitochondrially encoded cytochrome c oxidase subunit 2) is a critical component of complex IV in the mitochondrial respiratory chain. In canines, as in other mammals, this protein plays an essential role in cellular energy production. MT-CO2 functions primarily to transfer electrons from cytochrome c via its binuclear copper A center to the bimetallic center of the catalytic subunit 1 . As part of cytochrome c oxidase, it catalyzes the reduction of oxygen to water, the final step in the electron transport chain, enabling ATP synthesis through oxidative phosphorylation.
Unlike nuclear-encoded proteins, MT-CO2 is encoded by mitochondrial DNA, which has important implications for inheritance patterns, evolutionary studies, and genetic associations with performance traits in canids. The protein functions as a multi-pass membrane protein embedded in the mitochondrial inner membrane .
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What expression systems are most effective for producing functional recombinant canine MT-CO2?
Several expression systems can be employed for recombinant MT-CO2 production, each with specific considerations:
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Bacterial systems (E. coli): Require codon optimization of the canine MT-CO2 sequence and often result in inclusion bodies that need refolding. Using specialized strains like SHuffle or Origami can improve proper disulfide bond formation.
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Yeast systems: Pichia pastoris or Saccharomyces cerevisiae provide eukaryotic processing capabilities and have been successful for expressing mitochondrial membrane proteins similar to those studied in canids .
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Insect cell systems: Baculovirus expression systems offer better post-translational modifications and membrane protein folding.
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Mammalian cell systems: Provide the most native-like environment but with lower yields.
Regardless of the system chosen, several methodological considerations are crucial:
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Supplementing growth media with copper to facilitate proper cofactor incorporation
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Including solubility-enhancing fusion tags (SUMO, MBP, or thioredoxin)
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Optimizing induction conditions (temperature, inducer concentration, duration)
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Using appropriate detergents for extraction and purification
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How can researchers verify the structural integrity and functionality of recombinant canine MT-CO2?
A multi-technique approach is necessary to comprehensively characterize recombinant MT-CO2:
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SDS-PAGE and Western blotting: For assessing purity and confirming identity using MT-CO2-specific antibodies
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Mass spectrometry: For accurate molecular weight determination (expected ~28 kDa for canine MT-CO2) and peptide mapping
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Circular dichroism spectroscopy: To evaluate secondary structure elements characteristic of properly folded MT-CO2
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UV-visible spectroscopy: To verify copper center formation, which shows characteristic absorption features
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Electron transfer assays: To confirm functional capability using cytochrome c and oxygen consumption measurements
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Thermal stability assays: Such as differential scanning fluorimetry to assess protein stability
The presence of the copper cofactor is particularly critical for functional integrity and can be verified using electron paramagnetic resonance (EPR) spectroscopy.
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What polymorphisms have been identified in canine MT-CO2 and how might they be relevant to research?
Research in canids has identified several polymorphisms in MT-CO2. While specific data on domestic dogs is limited in the provided search results, studies in raccoon dogs identified six polymorphisms in the MT-CO2 gene that grouped into two distinct haplotypes (A and B) . All polymorphisms were synonymous (not changing amino acid sequence).
Methodological approaches to identify and characterize polymorphisms include:
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PCR amplification and direct sequencing of the MT-CO2 gene from diverse canine populations
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Bioinformatic analysis to classify variants into haplotypes
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Comparative sequence analysis across different canid species
Interestingly, despite being synonymous, these MT-CO2 polymorphisms showed significant associations with performance traits such as body weight, body size, and color type in raccoon dogs . This suggests that even synonymous mutations in MT-CO2 may have functional consequences in canids, potentially through effects on mRNA stability, translation efficiency, or interactions with nuclear-encoded proteins.
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