Recombinant Mycoplasma genitalium ATP synthase subunit c (atpE)
Description
Overview of Recombinant Mycoplasma genitalium ATP Synthase Subunit c (atpE)
Recombinant Mycoplasma genitalium ATP synthase subunit c (atpE) is a bioengineered protein derived from the F₀ sector of the ATP synthase complex. It plays a critical role in proton translocation across mitochondrial membranes in eukaryotes and bacterial bioenergetic processes. In M. genitalium, this protein is essential for ATP production via glycolysis, as the bacterium lacks respiratory chains and relies solely on fermentation for energy .
The recombinant form is produced in E. coli with a His-tag for purification, spanning the full-length sequence (1–102 amino acids) . While structural data for M. genitalium atpE is limited, studies on homologous proteins (e.g., human and yeast subunit c) reveal a conserved cylindrical structure critical for proton pumping .
Expression and Refolding
The recombinant atpE is expressed in E. coli under optimized conditions for solubility. Key steps include:
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Cloning: Insertion of the atpE gene into a plasmid vector (e.g., pET series) with a His-tag.
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Induction: IPTG-induced overexpression at 16–20°C to reduce inclusion body formation .
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Purification:
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Ni²⁺ affinity chromatography: Binds His-tagged protein.
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Size-exclusion chromatography: Ensures monodispersity.
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Quality Control
| Parameter | Specification | Method |
|---|---|---|
| Purity | >90% (SDS-PAGE) | Coomassie staining |
| Concentration | 0.1–1.0 mg/mL (reconstituted in deionized water) | Bradford assay |
| Storage | -20°C/-80°C (lyophilized powder) | Aliquoting recommended |
Data from commercial suppliers indicate high yield and stability, making it suitable for structural studies or functional assays .
Functional Studies
In M. genitalium, ATP synthase is critical for survival due to its reliance on glycolysis . Recombinant atpE could elucidate:
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Proton-translocating efficiency: Mutagenesis studies to map critical residues.
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Drug targets: Screening inhibitors of ATP synthase to combat antibiotic-resistant strains.
Comparative Insights
Challenges and Future Directions
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Limited Functional Data: Most studies focus on eukaryotic or other bacterial systems (e.g., E. coli, Enterococcus).
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Antigenic Potential: M. genitalium’s atpE may evade host immunity due to low homology with human proteins, making it a candidate for vaccine development .
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Therapeutic Targeting: Inhibiting ATP synthase could disrupt M. genitalium’s energy metabolism, but requires further validation .
Product Specs
Note: We prioritize shipping the format currently in stock. However, if you have specific format requirements, please indicate them during order placement. We will fulfill your request to the best of our ability.
Note: All our proteins are shipped with standard blue ice packs by default. If you require dry ice shipping, please inform us in advance. Additional fees may apply.
Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.
Target Background
KEGG: mge:MG_404
STRING: 243273.MgenG_010200000470
