Recombinant Solanum lycopersicum ATP synthase subunit a, chloroplastic (atpI)
Description
Definition and Production
Recombinant AtpI is produced by cloning the atpI gene from tomato (Solanum lycopersicum) into bacterial systems (e.g., E. coli) for large-scale purification. Key specifications include:
This protein is commercially available from suppliers like Creative BioMart and MyBioSource for research applications .
Domain Architecture
AtpI is a key component of the proton channel in the F₀ sector of cpATPase. It facilitates proton translocation across the thylakoid membrane, enabling ATP synthesis .
Key Features
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Proton Translocation: Critical for maintaining the proton gradient required for ATP generation .
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Interaction Networks: Binds with other cpATPase subunits (e.g., AtpB, AtpE) and assembly factors (e.g., BFA1, BFA3) .
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Redox Regulation: Modulates reactive oxygen species (ROS) homeostasis under stress .
Stress Response and Disease Resistance
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Pathogen Resistance: Overexpression of atpA (a related cpATPase subunit) in transgenic tobacco reduced Botrytis cinerea infection by enhancing ATP content (3.7–4.4-fold) and balancing ROS .
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Salt Tolerance: Transgenic plants overexpressing atpA exhibited improved salinity tolerance through enhanced ATPase activity and H⁺/Ca²⁺ efflux .
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Low-Temperature Adaptation: Overexpression of the ATP synthase β subunit in tomato increased cold tolerance by elevating antioxidant enzyme activity (SOD, POD, CAT) and photosynthetic efficiency .
Chloroplast Biogenesis
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AtpI interacts with assembly factors like AtpF and AtpG, which stabilize mRNA transcripts (e.g., atpE) during cpATPase assembly .
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Silencing atpE or atpF in cotton increased ROS levels, linking cpATPase subunits to redox homeostasis .
Agricultural Engineering
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Stress-Resilient Crops: Overexpression of cpATPase subunits enhances resistance to biotic (fungal pathogens) and abiotic (salinity, cold) stresses .
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Photosynthetic Optimization: Targeting AtpI could improve ATP synthesis efficiency, benefiting crop yields under suboptimal conditions .
Comparative Analysis
| Parameter | S. lycopersicum AtpI | Homolog in Acorus americanus |
|---|---|---|
| Amino Acid Sequence | 247 residues | 247 residues |
| Expression System | E. coli | E. coli |
| Key Function | Proton channel | Proton channel |
Future Directions
Product Specs
Note: We prioritize shipping the format currently in stock. However, if you have a specific format requirement, please indicate it when placing your order, and we will accommodate your request.
Note: All of our proteins are shipped with standard blue ice packs by default. If you require dry ice shipping, please inform us in advance, as additional fees will apply.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.
Target Background
KEGG: sly:3950464
Q&A
Basic Research Questions
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How does the expression of ATP synthase subunit a change under different stress conditions in tomato?
The expression of ATP synthase components, including subunit a, demonstrates significant plasticity in response to various stresses:
Under low-temperature stress:
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Expression of ATP synthase subunits is generally downregulated in wild-type tomato plants
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In transgenic plants overexpressing the ATP synthase β subunit gene, the photosynthetic apparatus maintains higher functionality during cold stress
Under pathogen infection:
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Expression of the ATP synthase CF1 α subunit (AtpA) is upregulated in response to fungal pathogens like Botrytis cinerea and beneficial fungi like Clonostachys rosea
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This upregulation is part of the plant's defense response mechanism
During fruit ripening:
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The expression of specific ATP synthase subunits changes, particularly with the incorporation of an atypical γ-subunit in chromoplasts that replaces the standard chloroplast γ-subunit
These expression changes reflect the adaptive responses of the tomato energy metabolism system to various environmental challenges.
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What techniques are most effective for isolating and purifying recombinant ATP synthase subunit a from tomato?
The isolation and purification of recombinant ATP synthase subunit a from tomato involves several specialized techniques:
Expression System Selection:
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E. coli expression systems are commonly used for producing recombinant ATP synthase subunits with N-terminal His tags for easier purification
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Agrobacterium-mediated transformation has been successfully used for generating transgenic tomato plants with modified ATP synthase expression
Organelle Isolation Protocol:
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Dark adaptation of plants (27 hours) to reduce starch accumulation
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High-yielding chloroplast isolation using optimized buffers for tomato leaves
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Chloroplast lysis and stromal protein collection by centrifugation
Protein Purification Steps:
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Affinity chromatography using His-tag binding for recombinant proteins
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Storage in appropriate buffer systems (typically Tris-based buffer with 50% glycerol)
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Lyophilization into powder form for long-term storage
Quality Assessment:
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SDS-PAGE for purity determination (>90% purity standard)
For researchers working with tomato chloroplast proteins, a high-yielding protocol has been developed that allows the isolation of highly pure chloroplast and stromal proteins from tomato leaves, yielding comprehensive identification of over 1,278 stromal proteins .
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