Recombinant Glycine max ATP synthase subunit c, chloroplastic (atpH)
Description
Recombinant Production and Purification
Expression Systems:
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Bacterial Systems: E. coli is commonly used for high-yield production. The spinach homolog (c₁ subunit) was expressed in E. coli and purified via maltose-binding protein (MBP) fusion tags, enabling isolation of mg quantities .
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Cell-Free Systems: Alternative approaches include cell-free expression for structural studies .
Purification Methods:
| Step | Method |
|---|---|
| Tagging | N-terminal His-tag for immobilized metal affinity chromatography (IMAC) . |
| Buffer Optimization | Tris-based buffer with 50% glycerol for stability; storage at -20°C or -80°C . |
Challenges:
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Membrane Integration: Subunit c’s hydrophobic nature complicates solubility. Spinach c₁ required refolding from inclusion bodies .
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Stoichiometry: Variations in c-subunit ring size (cₙ) influence proton-to-ATP ratios; recombinant systems enable controlled studies .
Research Applications and Functional Insights
Role in ATP Synthase Assembly:
Subunit c interacts with other F₀ components (e.g., subunits a, b, I) and CF₁ subunits (α, β, γ, ε, δ) to form the functional ATP synthase holoenzyme . Mutants lacking proper RNA processing (e.g., bfa2 in Arabidopsis) show reduced atpH transcript stability, impairing subunit c accumulation and ATP synthase activity .
Interactions with Regulatory Proteins:
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BFA1: A nucleus-encoded chaperone that facilitates CF₁ assembly by interacting with β and γ subunits, indirectly affecting subunit c integration .
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BFA2: A PPR protein essential for stabilizing atpH/F/A transcripts; its loss reduces ATP synthase levels by ~75% .
Functional Studies:
Product Specs
Note: We will prioritize shipping the format currently in stock. However, if you have specific format requirements, please indicate them in your order remarks. We will prepare the product according to your specifications.
Note: All our proteins are shipped with standard blue ice packs. 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: gmx:3989296
