Recombinant Leptospira biflexa serovar Patoc ATP synthase subunit delta (atpH)
Description
Overview of Recombinant Leptospira biflexa serovar Patoc ATP Synthase Subunit Delta (atpH)
Recombinant ATP synthase subunit delta (atpH) from Leptospira biflexa serovar Patoc is a bioengineered protein derived from the ATP synthase complex, a key enzyme in bacterial energy production. ATP synthase is responsible for converting chemical energy into ATP through oxidative phosphorylation, with subunit delta (atpH) playing a structural role in the F1 sector, facilitating proton translocation and ATP synthesis. While no specific studies directly address the atpH subunit in L. biflexa, its function aligns with conserved mechanisms observed in other bacterial species .
Recombinant Production Methods
Recombinant atpH production involves heterologous expression in surrogate hosts like E. coli or L. biflexa itself. Key steps include:
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Cloning: Amplification of the atpH gene from L. biflexa genomic DNA using primers designed for restriction sites (e.g., EcoRI/XhoI) .
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Expression: Transformation into expression vectors (e.g., pET28a) under inducible promoters (e.g., T7 lac) .
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Purification: Affinity chromatography (Ni-NTA or GST tags) followed by size-exclusion chromatography .
| Host System | Advantages | Challenges |
|---|---|---|
| E. coli | High yield, cost-effective | Possible misfolding due to lack of native chaperones |
| L. biflexa | Native posttranslational modifications | Slower growth, lower yields |
Biochemical and Functional Characterization
Functional assays for recombinant atpH include:
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Proton channel activity: Measured using F0 sector reconstitution assays .
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ATP synthesis: Coupled with F1 sector subunits to assess catalytic efficiency .
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Structural studies: X-ray crystallography or cryo-EM to resolve interactions with F0 subunits (e.g., atpB, atpE) .
Relevance to Leptospira Pathogenesis
While L. biflexa is non-pathogenic, studying its ATP synthase may inform mechanisms of energy metabolism in pathogenic Leptospira spp. (e.g., L. interrogans), which require ATP synthase for survival in hosts .
Applications in Research
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Vaccine Development: Subunit vaccines targeting ATP synthase components could disrupt energy production in pathogens .
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Antibiotic Discovery: Inhibitors of ATP synthase (e.g., bedaquiline) represent novel targets for leptospirosis treatment .
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Structural Biology: Insights into atpH interactions may guide rational drug design .
Challenges and Future Directions
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Limited functional data: No direct studies on L. biflexa atpH hinder mechanistic understanding .
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Expression complexity: Native folding requires co-expression with chaperones (e.g., GroEL-GroES) .
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Pathogenic relevance: Confirming atpH’s role in L. interrogans pathogenesis requires comparative studies .
Product Specs
Target Background
KEGG: lbi:LEPBI_I0804
STRING: 456481.LEPBI_I0804
