Recombinant Brucella canis NADH-quinone oxidoreductase subunit K (nuoK)
Description
Functional Residues
While specific mutational studies on Brucella canis nuoK are absent, conserved residues in homologous proteins (e.g., Mycobacterium bovis nuoK , E. coli NDH-1 ) highlight critical regions:
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Glu-36 (TM2): Essential for proton translocation; mutation to Ala abolishes NDH-1 activity .
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Glu-72 (TM3): Modulates activity; substitutions reduce proton pumping efficiency .
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Cytoplasmic Loop (Arg-25, Arg-26, Asn-27): Vital for energy coupling .
Bioenergetic Contributions
The NDH-1 complex, including nuoK, catalyzes electron transfer from NADH to quinone while pumping protons across the membrane. In Brucella, this process is likely critical for intracellular survival in host cells, where ATP production and redox balance are essential.
Pathogenic Relevance
While nuoK itself is not directly targeted in current diagnostic assays for B. canis (unlike Omp25 or PdhB/Tuf ), its role in maintaining membrane potential and ATP synthesis positions it as a potential therapeutic target.
Expression and Purification Workflow
Recombinant nuoK is typically produced via:
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Cloning: Insertion of the nuoK gene into a plasmid vector (e.g., pET series).
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Expression: Induction in E. coli with IPTG, followed by cell lysis.
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Purification: Affinity chromatography via His-tag, followed by size-exclusion or ion-exchange chromatography.
| Parameter | Optimal Condition |
|---|---|
| Reconstitution | Deionized sterile water (0.1–1.0 mg/mL) |
| Glycerol Addition | 5–50% (50% default for long-term storage) |
| Storage | -20°C/-80°C; avoid freeze-thaw cycles |
Source: Chemicalbook production guidelines
Diagnostic Potential
While nuoK is not currently used in serological assays (e.g., ELISA for B. canis targets PdhB or Tuf ), its conserved residues could be explored as epitopes for vaccine development.
Research Gaps
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Functional Characterization: Direct studies on B. canis nuoK’s role in pathogenesis are lacking.
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Antigenicity: No data exist on nuoK’s immunogenicity in canine or human sera.
Opportunities
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Therapeutic Targets: Inhibitors targeting nuoK’s proton-translocation residues could disrupt B. canis bioenergetics.
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Structural Analysis: Cryo-EM or X-ray crystallography of B. canis NDH-1 could elucidate species-specific interactions.
Product Specs
Note: While we will prioritize shipping the format currently in stock, we understand your specific needs. Should you require a particular format, please indicate your preference in your order notes, and we will fulfill your request whenever possible.
Note: Our standard shipping method includes normal blue ice packs. For shipments with dry ice, please contact us in advance to arrange for the additional service and associated charges.
Generally, liquid forms have a shelf life of 6 months at -20°C/-80°C. Lyophilized forms typically have a shelf life of 12 months at -20°C/-80°C.
Target Background
KEGG: bcs:BCAN_A0827
