Recombinant Mesoplasma florum ATP synthase subunit c (atpE)
Description
Overview of Recombinant Mesoplasma florum ATP Synthase Subunit c (atpE)
Recombinant Mesoplasma florum ATP synthase subunit c (atpE) is a heterologously expressed protein derived from the atpE gene of Mesoplasma florum, a mollicute bacterium. This subunit is a critical component of the F₀ sector of the F₀F₁ ATP synthase, a transmembrane enzyme responsible for converting electrochemical proton gradients into ATP. The recombinant form is engineered for structural, functional, and immunological studies, often produced in Escherichia coli systems with affinity tags for purification .
2.1. Amino Acid Sequence and Domain Architecture
The recombinant atpE protein spans 104 amino acids (full-length sequence:
MLFTDYMANFLVGYFSVLSSIMPLLAETSSTGEGLKLLGAGVAIIGVAGAGIGQGAVGQG ACMAIGRNPEMAPKITSTMIIAAGIAESGAIYALVVAILLIFVA), with a predicted α-helical secondary structure . The N-terminal 10×His-tag facilitates purification via nickel affinity chromatography .
2.2. Role in ATP Synthase Mechanism
As part of the F₀ sector, atpE contributes to the proton-conductive c-ring, which drives rotation of the F₁ stalk during ATP synthesis . In mollicutes like Mesoplasma florum, ATP synthase operates primarily in ATP hydrolysis mode to maintain transmembrane proton gradients .
3.1. Expression System
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Vector: Modified pMAL-c2x plasmid fused with atpE for high-yield expression .
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Chaperones: Co-expression with DnaK, DnaJ, and GrpE enhances solubility .
3.2. Purification Protocol
4.2. Functional Characterization
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Enzymatic Activity: Used to reconstitute active ATP synthase complexes in vitro .
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Immunological Studies: Serves as an antigen in ELISA kits for detecting atpE-specific antibodies .
4.3. Applications in Molecular Biology
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Protein-Protein Interactions: Investigating interactions with F₀ subunits (e.g., a, b, d) or F₁ components .
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Drug Targeting: Screening inhibitors of ATP synthase in bacterial pathogens .
Comparative Analysis of ATP Synthase Subunits
| Feature | Mesoplasma florum atpE (F₀) | Spinach Chloroplast atpH (F₀) |
|---|---|---|
| Gene Origin | Nuclear (recombinant) | Plastid-encoded |
| Tag | 10×His | MBP (maltose-binding protein) |
| Primary Use | Structural studies, ELISA | Reconstituting c-rings |
| Expression System | E. coli | E. coli |
Limitations and Challenges
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Low Functional Redundancy: Unlike mammalian subunit c isoforms (P1/P2/P3), Mesoplasma florum atpE lacks alternative splicing variants .
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Biochemical Complexity: Purification requires optimization for membrane protein solubility .
References
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Dyer, M. R., et al. (1989). "Subunit c isoforms in mammalian ATP synthase." J. Biol. Chem.
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Alavian, K. N., et al. (2014). "c-Subunit leak regulates mitochondrial ATP synthase activity." PNAS
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Cusabio. (2023). "Recombinant Mesoplasma florum ATP synthase subunit c (atpE)." Product Datasheet.
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Dieckmann, C. L., et al. (2010). "Recombinant production of spinach chloroplast ATP synthase subunit c." PLoS ONE.
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American Science. (2023). "ELISA Recombinant Mesoplasma florum ATP synthase subunit c (atpE)." Product Page.
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Ye, F., et al. (2010). "Spiroplasma citri genome sequence reveals ATP synthase operon." Appl. Environ. Microbiol.
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Béven, L., et al. (2012). "F₁-like ATPases in mycoplasmas." PLoS ONE.
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Lamontagne Boulet, M., et al. (2018). "ATP synthase as a target for bactericidal agents." Antimicrob. Agents Chemother.
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Wikipedia. (2025). "ATP synthase." Structure and Function.
Product Specs
Note: We will prioritize shipping the format we have in stock. However, if you have specific requirements for the format, please indicate them in your order. We will fulfill your request if possible.
Note: All our proteins are shipped with normal blue ice packs by default. If you require dry ice shipping, please inform us in advance as additional charges 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: mfl:Mfl110
STRING: 265311.Mfl110
