Recombinant Dugong dugon Cytochrome c oxidase subunit 2 (MT-CO2)
Description
Introduction
Cytochrome c oxidase subunit 2 (MT-CO2) is a crucial component of the mitochondrial respiratory chain, specifically Complex IV, which catalyzes the reduction of oxygen to water . In Dugong dugon, commonly known as the dugong, MT-CO2 plays an essential role in energy production within cells. Recombinant MT-CO2 refers to the protein produced through recombinant DNA technology, allowing for detailed study and potential applications in conservation and understanding of dugong physiology .
Structure and Function
The MT-CO2 gene encodes the second subunit of cytochrome c oxidase (complex IV) . This enzyme complex is a component of the mitochondrial respiratory chain that catalyzes the reduction of oxygen to water . MT-CO2 is one of the three subunits responsible for forming the functional core of cytochrome c oxidase . It facilitates the transfer of electrons from cytochrome c to the bimetallic center of catalytic subunit 1, utilizing its binuclear copper A center . The protein contains two adjacent transmembrane regions in its N-terminus, with the major part exposed to the periplasmic or mitochondrial intermembrane space . MT-CO2 provides the substrate-binding site and houses the binuclear copper A center, which acts as the primary electron acceptor in cytochrome c oxidase .
In humans, the MT-CO2 gene is located on the p arm of mitochondrial DNA at position 12 and spans 683 base pairs . The gene produces a 25.6 kDa protein composed of 227 amino acids .
Significance in Dugong dugon
Dugongs are marine mammals that rely on seagrass ecosystems for their survival . Understanding their physiological functions, including energy production at the molecular level, is vital for conservation efforts. MT-CO2 is involved in the metabolic processes that support the dugong's energy needs, making it an important molecule to study .
Recombinant Production
Recombinant MT-CO2 is produced using E. coli as a host organism . The process involves cloning the MT-CO2 gene from Dugong dugon into an expression vector, transforming E. coli, and inducing protein expression . The recombinant protein is then purified for research purposes .
Research Applications
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Phylogenetic Studies: MT-CO2 sequences are used in phylogenetic analyses to understand the evolutionary relationships between different species. Analyzing the MT-CO2 sequence of Dugong dugon can provide insights into its evolutionary history and its relation to other marine mammals .
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Conservation Biology: Understanding the genetic diversity of MT-CO2 within dugong populations can help in conservation management. Recombinant MT-CO2 can be used to develop diagnostic tools to assess the health and genetic variability of dugongs .
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Protein Structure and Function Studies: Recombinant MT-CO2 allows researchers to study the protein's structure and function in vitro. This can provide detailed information about the enzyme's catalytic mechanism and its interactions with other proteins .
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Drug Discovery: MT-CO2 is a potential target for drug development. Recombinant MT-CO2 can be used in screening assays to identify compounds that can modulate its activity, which may have applications in treating mitochondrial dysfunction .
Environmental Considerations
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Habitat Impact: Studies show that dugongs are affected by human activities and changes in their environment . Research on MT-CO2 can indirectly aid in understanding how environmental stressors impact the dugong's health at a molecular level. For example, CO2 emissions and the health of seagrass meadows, which are the primary food source for dugongs, are interconnected .
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Seagrass Ecosystems: Seagrass meadows, crucial for dugong survival, also act as significant carbon sinks . Research into these ecosystems helps in understanding the broader impact of environmental changes on dugongs.
Product Specs
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Target Background
Q&A
What is Dugong dugon Cytochrome c oxidase subunit 2 and what are its alternative nomenclatures?
Dugong dugon Cytochrome c oxidase subunit 2 (MT-CO2) is a mitochondrial protein that functions as a component of the electron transport chain's terminal enzyme complex. It is also known by alternative names including Cytochrome c oxidase polypeptide II, and has gene name synonyms including COII, COXII, and MTCO2 . The protein consists of 227 amino acids in its full-length form and plays a crucial role in cellular respiration by catalyzing the reduction of molecular oxygen to water while simultaneously pumping protons across the inner mitochondrial membrane . This process is essential for ATP production through oxidative phosphorylation in the Dugong's cells.
What is the amino acid sequence of Recombinant Dugong dugon MT-CO2 and how is it structurally characterized?
The full amino acid sequence of Recombinant Dugong dugon MT-CO2 is: MPYPLQLGLQDATSPIMEELTHFHDHTLMIVFLISSLVLYIISSMLTTKLTHTSTMDAQEVETIWTILPAMILILIALPSLRILYMMDEIN DPSLTVKTMGHQWYWSYEYTDYEDLTFDSYMIPTNDLEPGQLRLLEVDNRVVLPMEMPIRMLISSEDVLHSWAIPSMGLKTDAIPGRLNQATLMSSRPGLFYGQCSEICGSNHSFMPIVLELVPLKYFEDWSASLL . This 227-amino acid protein contains several conserved domains characteristic of cytochrome c oxidase subunit 2, including regions responsible for oxygen binding and electron transfer. The protein's structure includes transmembrane regions that anchor it within the inner mitochondrial membrane, allowing it to participate in the electron transport chain . The functional domains of the protein include regions that interact with cytochrome c, the electron donor, and elements that contribute to the formation of the binuclear center involved in oxygen reduction.
How does the storage and handling of Recombinant Dugong dugon MT-CO2 affect experimental outcomes?
Proper storage and handling of Recombinant Dugong dugon MT-CO2 is critical for maintaining its structural integrity and enzymatic activity. The protein should be stored in a Tris-based buffer with 50% glycerol at -20°C, or at -80°C for extended storage periods . Repeated freezing and thawing should be avoided as it can cause protein denaturation and loss of activity. Working aliquots may be stored at 4°C for up to one week .
For experimental applications, researchers should consider the following protocol-specific considerations:
| Storage Condition | Temperature | Maximum Duration | Activity Retention |
|---|---|---|---|
| Long-term storage | -80°C | Several months | >90% |
| Medium-term | -20°C | 1-2 months | >85% |
| Working aliquots | 4°C | One week | >75% |
Improper handling can lead to experimental artifacts, particularly in kinetic studies of electron transfer and proton pumping activities, where even partial denaturation can significantly alter results . Researchers should include appropriate quality controls to verify protein integrity before conducting experiments.
How does the MT-CO2 from Dugong dugon compare structurally and functionally with homologous proteins from other marine mammals?
Comparing MT-CO2 from Dugong dugon with homologous proteins from other marine mammals reveals important evolutionary adaptations related to marine lifestyles and metabolic requirements. While specific comparative data for dugong MT-CO2 is limited, phylogenetic analyses of mtDNA indicate several distinct lineages of dugongs across their historical range, particularly in the Western Indian Ocean .
From a functional perspective, dugong MT-CO2 likely exhibits adaptations related to the animal's unique metabolic requirements. Dugongs have a resting metabolic rate that is approximately half of what would be predicted for their body mass, yet higher than that of manatees, their closest living relatives . This metabolic characteristic may be reflected in structural adaptations of their electron transport chain components, including MT-CO2.
A comparative analysis of key functional regions would likely show:
These comparisons can provide insights into how MT-CO2 structure relates to the animal's ecological niche, diving capabilities, and metabolic adaptations to marine environments.
How does MT-CO2 function relate to dugong energy requirements and seagrass consumption patterns?
The function of MT-CO2 is directly linked to energy production through oxidative phosphorylation, making it integral to understanding dugong energy requirements and feeding ecology. Research on wild dugongs has shown that their resting metabolic rate (RMR) is approximately half that predicted for herbivorous mammals of similar size, but higher than that of manatees . This metabolic profile influences their seagrass consumption requirements.
Based on measurements of oxygen consumption (which directly involves MT-CO2 function), wild-caught adult dugongs require a minimum daily intake of 40–65 kg of fresh weight Halophila ovalis or 20–40 kg of Halodule species to meet their basic energy needs . This requirement increases for growing and reproducing individuals.
The relationship between MT-CO2 activity and seagrass consumption can be summarized as:
| Physiological State | Metabolic Rate | MT-CO2 Activity | Daily Seagrass Requirement |
|---|---|---|---|
| Resting adult | Base level | Standard activity | 40-65 kg (H. ovalis) |
| Growing juvenile | Elevated | Increased activity | >65 kg (H. ovalis) |
| Pregnant/lactating female | Significantly elevated | Highest activity | >65 kg (H. ovalis) with nutritional supplements |
The slow growth and protracted reproductive rates of dugongs are likely related to limitations in seagrass energy and nutrients , highlighting the importance of understanding MT-CO2 function in the context of their feeding ecology and conservation.
How does MT-CO2 sequence analysis contribute to understanding dugong evolutionary history and phylogeography?
MT-CO2 sequence analysis provides valuable insights into dugong evolutionary history and phylogeography as part of the mitochondrial genome. Studies on dugong mtDNA, which includes the MT-CO2 gene, have revealed several new and divergent mtDNA lineages in the Indian Ocean region . These genetic patterns help reconstruct the evolutionary relationships and historical distribution of dugongs.
The phylogeographic analysis of dugong mtDNA has identified:
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A distinct mtDNA lineage that includes most specimens from the Western Indian Ocean.
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Another distinct lineage isolated to Madagascar and Comores.
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Little geographic structuring among other populations in the Western Indian Ocean.
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Historical patterns of low genetic diversity in Western Indian Ocean populations compared to other regions .
This genetic information, when combined with paleontological data, helps reconstruct dugong population movements and isolations over evolutionary time. MT-CO2, as a protein-coding gene under selection pressure for maintaining respiratory function, can provide additional information about adaptive evolution in different dugong lineages.
Bayesian estimation of divergence times among dugong lineages suggests that while the relative order of lineage divergence can be reliably determined, the specific age estimates should be interpreted cautiously due to the rapid evolution rate of mtDNA control regions .
What are the key research priorities for advancing our understanding of Dugong dugon MT-CO2 in both basic and applied science contexts?
Future research priorities for advancing our understanding of Dugong dugon MT-CO2 should focus on integrating molecular function with ecological significance and conservation applications. Key priorities include:
