Recombinant Salmonella enteritidis PT4 ATP synthase subunit c (atpE)

Shipped with Ice Packs
In Stock

Description

Vaccine Development

The atpE protein is highlighted as a potential vaccine candidate due to its conserved role in bacterial survival. Salmonella ATP synthase subunit c is critical for energy production, making it a viable target for disrupting pathogenic processes. Suppliers note its use in preclinical studies to elicit immune responses, though specific efficacy data remain unpublished .

ELISA and Serological Studies

Recombinant atpE serves as an antigen in enzyme-linked immunosorbent assays (ELISA) to detect anti-Salmonella antibodies. Its high purity ensures minimal cross-reactivity, enabling precise serological analysis .

Functional Insights from Related Research

Although no direct studies on S. enteritidis atpE are available, broader research on bacterial ATP synthases provides context:

  • Energy Metabolism: ATP synthase subunit c facilitates proton translocation, driving ATP synthesis. In Salmonella Pullorum, energy metabolism pathways (e.g., hydrogenases, succinate dehydrogenase) are downregulated under stress, highlighting the enzyme’s role in survival .

  • Virulence and Host Interaction: While atpE itself is not directly linked to virulence, ATP synthase activity supports intracellular replication and evasion of host immune responses. For example, Salmonella mutants deficient in energy metabolism genes exhibit reduced fitness in macrophages .

Product Specs

Form
Lyophilized powder
Note: We prioritize shipping the format currently in stock. However, if you have a specific format requirement, please indicate it when placing your order. We will accommodate your request to the best of our ability.
Lead Time
Delivery time may vary depending on the purchasing method and location. For precise delivery time information, please consult your local distributor.
Note: All our proteins are shipped with standard blue ice packs. If you require dry ice shipment, please inform us beforehand as additional charges may apply.
Notes
Repeated freezing and thawing is not recommended. For short-term storage, store working aliquots at 4°C for up to one week.
Reconstitution
It is recommended to briefly centrifuge the vial before opening to ensure the contents are at the bottom. Reconstitute the protein with deionized sterile water to a concentration of 0.1-1.0 mg/mL. We recommend adding 5-50% glycerol (final concentration) and aliquoting for long-term storage at -20°C/-80°C. Our standard final glycerol concentration is 50%. Customers can use this as a reference.
Shelf Life
Shelf life is influenced by various factors including storage conditions, buffer components, temperature, and the inherent stability of the protein.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. Lyophilized form has a shelf life of 12 months at -20°C/-80°C.
Storage Condition
Upon receipt, store at -20°C/-80°C. Aliquot for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
Tag type is determined during the manufacturing process.
The tag type will be determined during production. If you have a specific tag type preference, please inform us, and we will prioritize developing the specified tag.
Synonyms
atpE; SEN3684; ATP synthase subunit c; ATP synthase F(0 sector subunit c; F-type ATPase subunit c; F-ATPase subunit c; Lipid-binding protein
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-79
Protein Length
full length protein
Species
Salmonella enteritidis PT4 (strain P125109)
Target Names
atpE
Target Protein Sequence
MENLNMDLLYMAAAVMMGLAAIGAAIGIGILGGKFLEGAARQPDLIPLLRTQFFIVMGLV DAIPMIAVGLGLYVMFAVA
Uniprot No.

Target Background

Function
F(1)F(0) ATP synthase catalyzes the production of ATP from ADP in the presence of a proton or sodium gradient. F-type ATPases comprise two structural domains: F(1), the extramembraneous catalytic core, and F(0), the membrane proton channel, connected via a central stalk and a peripheral stalk. During catalysis, ATP synthesis within the catalytic domain of F(1) is coupled to proton translocation through a rotary mechanism involving the central stalk subunits. This subunit is a key component of the F(0) channel and directly participates in membrane translocation. A homomeric c-ring consisting of 10-14 subunits forms the central stalk rotor element, interacting with the F(1) delta and epsilon subunits.
Database Links

KEGG: set:SEN3684

Protein Families
ATPase C chain family
Subcellular Location
Cell inner membrane; Multi-pass membrane protein.

Q&A

What is ATP synthase subunit c (atpE) and its role in Salmonella enteritidis PT4?

ATP synthase subunit c is a building block of the membrane rotor within the F₀ complex of ATP synthase. It functions as part of the molecular turbine that uses transmembrane proton flow to generate ATP. In Salmonella enteritidis PT4, this protein forms a ring structure (c-ring) that rotates as protons pass through the membrane, driving the conformational changes necessary for ATP synthesis . The atpE gene is highly conserved across Salmonella serovars, reflecting its essential function in cellular bioenergetics. The amino acid sequence of atpE (position 1-79) contains the critical residues involved in proton binding and translocation that power the rotational mechanism of ATP synthase .

How does subunit c interact with other components of the ATP synthase complex?

Subunit c interacts primarily with subunit a, which forms part of the membrane stator of the ATP synthase. This interaction is crucial for assembling the proton channel of ATP synthase. Research has demonstrated that subunit a can be molecularly fused with subunit c, and this fusion is capable of incorporating into the ATP synthase complex, providing a valuable structural model for studying the proton channel . The interaction between subunit a and monomeric subunit c is a key initial step in ATP synthase assembly, as it triggers the insertion of subunit a into the membrane and initiates formation of the a-c complex, which constitutes the ion-translocating module of ATP synthase . This interaction precedes the assembly of the c-ring, with the fused c subunit either incorporating into the c-ring or remaining on its periphery depending on the orientation of the transmembrane helices in the fusion protein .

What experimental approaches can effectively distinguish between functional and non-functional atpE mutations?

Researchers can distinguish between functional and non-functional atpE mutations through several methodological approaches:

  • Complementation studies: Wild-type and mutant forms of atpE can be expressed in atpE-deficient strains to determine if function is restored.

  • Proton translocation assays: Measuring pH changes across membranes containing reconstituted ATP synthase complexes with different atpE variants.

  • ATP synthesis measurements: Quantifying ATP production in systems with various atpE mutations can directly assess functional impact.

  • Molecular fusion approaches: As demonstrated in research, creating fusion proteins between subunit a and subunit c can help determine if specific orientations and interactions are functional. For example, researchers have observed that a/c fusion proteins with correct orientation of transmembrane helices could be inserted into the membrane and co-incorporated into the F₀ complex, while fusions with incorrect orientation required wild-type subunit c for membrane insertion .

  • Motility assays: For Salmonella, which uses ATP synthase to power flagellar rotation, motility tests can indirectly assess atpE function, particularly in the context of type-III secretion systems which share evolutionary relationships with ATP synthase components .

How is the atpE gene organized within the ATP synthase operon of S. enteritidis PT4?

The atpE gene in Salmonella enteritidis PT4 is part of a larger atp operon that encodes multiple subunits of the ATP synthase complex. Analysis of the complete genome of S. enteritidis PT4 (strain P125109, EMBL accession no. AM933172) reveals high conservation of this core metabolic machinery . The gene organization follows the typical arrangement found in most bacteria, with atpE positioned within a cluster of genes encoding other ATP synthase components.

Comparative genome analysis between S. enteritidis PT4 and S. Typhimurium LT2 shows extremely high nucleotide identity (98.98%) between shared orthologs, suggesting the ATP synthase genes are part of the extensive core gene-set that maintains colinearity between these serovars . This high conservation reflects the essential nature of ATP synthase genes for bacterial viability and energy metabolism across Salmonella species.

What evolutionary insights can be drawn from comparing atpE across different Salmonella serovars?

Comparative genomic analysis reveals several important evolutionary insights about atpE:

  • High conservation: The atpE gene shows remarkable sequence conservation across Salmonella serovars, including between S. enteritidis PT4 and S. Typhimurium LT2, which share >90% of their coding sequences as part of an extensive core gene-set .

  • Evolutionary relationship with type-III secretion systems: Research has demonstrated a strong homology between cytoplasmic components of type-III secretion systems and the F₀F₁ ATP synthase. This suggests that the bacterial flagellum, which includes components analogous to ATP synthase, may have evolved from a proto F₀F₁-ATP synthase .

  • Evolutionary sequence: Studies indicate that a proto ATPase may have been added to a primordial proton-powered type-III export system, with the evolutionary benefit of facilitating the export process. This is supported by findings that ATPase activity can be dispensable for type-III protein export in Salmonella under conditions of increased proton motive force .

  • Minimal pseudogenization: Unlike many virulence-associated genes that undergo pseudogenization in host-adapted strains, core metabolic genes like atpE typically maintain their functionality. For example, while S. Gallinarum (a highly host-adapted chicken pathogen) harbors a significantly higher number of pseudogenes compared to S. enteritidis PT4, essential metabolic genes remain functional .

This evolutionary perspective provides important context for understanding the fundamental role of atpE in bacterial physiology and its relationship to other bacterial systems.

How can recombinant S. enteritidis PT4 atpE be utilized in vaccine development?

Recombinant S. enteritidis PT4 atpE protein has several potential applications in vaccine development:

  • As a component in attenuated live vaccines: Salmonella strains with modified atpE could serve as attenuated vaccine carriers. Research has demonstrated that Salmonella spp. can be genetically modified to create multivalent live carrier vaccines for simultaneous immunization against several unrelated pathogens .

  • Engineering acid resistance: The ATP synthase is linked to acid resistance mechanisms in Salmonella. Researchers have re-engineered acid resistance systems in Salmonella vaccine strains to enhance survival under acidic conditions, which could improve vaccine efficacy when administered orally . Similar approaches could potentially utilize atpE modifications.

  • Antigen presentation platform: As a component of the bacterial membrane, atpE could potentially be engineered to present foreign epitopes or antigens, leveraging Salmonella's ability to deliver effector proteins to host cells through type-III secretion systems .

  • Target for attenuating mutations: Introducing specific mutations in atpE that reduce ATP synthesis efficiency without abolishing it completely could create attenuated strains with reduced virulence but maintained immunogenicity.

When developing such applications, researchers must consider that recombinant Salmonella atpE protein products are strictly for research purposes and cannot be used directly on humans or animals without appropriate regulatory approval and clinical trials .

What purification methods are most effective for recombinant S. enteritidis PT4 atpE protein?

Purification of recombinant S. enteritidis PT4 atpE presents challenges due to its hydrophobic nature as a membrane protein. The following methodological approaches have proven effective:

  • Expression systems selection: Multiple expression systems can be utilized, including E. coli, yeast, baculovirus, or mammalian cells . E. coli systems are commonly preferred for initial screening due to their high yield and cost-effectiveness, though eukaryotic systems may provide better folding for structural studies.

  • Membrane protein extraction: Effective extraction requires careful solubilization using detergents such as n-dodecyl-β-D-maltoside (DDM) or digitonin that maintain protein structure while removing it from the lipid bilayer.

  • Affinity chromatography: His-tagged recombinant atpE can be purified using immobilized metal affinity chromatography (IMAC). For fusion constructs like the a/c fusion proteins described in the literature, appropriate tag placement is critical to ensure accessibility without interfering with structure .

  • Size exclusion chromatography: This technique helps separate monomeric atpE from aggregates or incomplete translation products, which is particularly important when studying the monomeric form's interaction with subunit a .

  • Reconstitution methods: For functional studies, purified atpE must often be reconstituted into liposomes or nanodiscs to restore its native membrane environment.

The choice of purification method should be guided by the intended experimental application, whether structural analysis, functional studies, or immunological investigation.

How does the proton motive force interact with atpE function in ATP synthesis?

The interaction between proton motive force and atpE function represents a fundamental aspect of bioenergetics:

  • Primary energy source: The proton motive force (PMF) serves as the primary energy source driving ATP synthesis through the ATP synthase complex. Protons flowing through the a/c interface of ATP synthase cause rotation of the c-ring (composed of multiple copies of subunit c/atpE) .

  • Dispensability of ATPase activity: Research has revealed that under conditions of increased proton motive force, the requirement for ATPase activity can be bypassed in certain secretion systems related to ATP synthase. This finding has important implications for understanding the evolutionary relationship between ATP synthase and type-III secretion systems .

  • Experimental evidence: Studies with Salmonella have demonstrated that mutations increasing the proton motive force allowed formation of functional flagella even in the absence of type-III ATPase activity. This was further supported by observations that increased proton motive force could bypass the requirement of the Salmonella pathogenicity island 1 virulence-associated type-III ATPase for secretion .

  • Functional relationship: The role of subunit c (atpE) in this process involves forming the ion-binding sites that capture and release protons during rotation, converting the energy of the proton gradient into mechanical motion that drives ATP synthesis. The specific amino acid residues within atpE that participate in proton binding are critical for this function .

This fundamental relationship between proton motive force and atpE function underscores the central role of this protein in cellular bioenergetics and bacterial physiology.

What structural features of atpE are critical for c-ring assembly and function?

Several structural features of atpE are essential for proper c-ring assembly and ATP synthase function:

Understanding these structural features has been enhanced through experimental approaches such as creating fusion proteins between subunits a and c, which have provided valuable insights into the organization of the proton channel and the assembly process of ATP synthase .

How does S. enteritidis PT4 atpE compare with atpE from other bacterial pathogens?

Comparative analysis of S. enteritidis PT4 atpE with homologs from other bacterial pathogens reveals important similarities and differences:

This high conservation of atpE across diverse bacterial species reflects its fundamental role in cellular bioenergetics. Comparative genomic analysis between S. enteritidis PT4 and S. Typhimurium LT2 shows extremely high nucleotide identity (98.98%) between shared orthologs , suggesting that ATP synthase genes like atpE are part of the extensive core gene-set that maintains evolutionary stability across Salmonella serovars.

What evidence supports the evolutionary relationship between ATP synthase and type-III secretion systems?

Several lines of evidence support an evolutionary relationship between ATP synthase (which includes atpE) and type-III secretion systems:

  • Structural homology: The cytoplasmic components of the type-III secretion system share strong homology with the F₀F₁ ATP synthase, suggesting a common evolutionary origin .

  • Functional parallels: Both systems are involved in energy-dependent transport across membranes - ATP synthase transports protons for energy production, while type-III secretion systems transport proteins across membranes.

  • Experimental evidence of functional overlap: Research has demonstrated that the flagellar type-III secretion apparatus utilizes both the energy of the proton motive force and ATP hydrolysis to energize substrate translocation, similar to how ATP synthase utilizes the proton motive force .

  • Dispensability of ATPase: Studies have shown that functional flagella can form in the absence of type-III ATPase activity when mutations increase the proton motive force and flagellar substrate levels. This suggests that the proton motive force is the more fundamental energy source, with ATPase activity serving an enhancing role that may have been added later in evolutionary history .

  • Evolutionary model: The evidence supports a model where "a proto ATPase was added to a primordial proton-powered type-III export system with the evolutionary benefit of facilitating the export process" . This indicates that the original export system relied solely on the proton motive force, with the ATPase component being a later evolutionary addition that enhanced efficiency.

This evolutionary relationship provides important context for understanding both ATP synthase function and the mechanisms of bacterial protein secretion systems that are critical for pathogenicity.

Quick Inquiry

Personal Email Detected
Please use an institutional or corporate email address for inquiries. Personal email accounts ( such as Gmail, Yahoo, and Outlook) are not accepted. *
© Copyright 2025 TheBiotek. All Rights Reserved.