RCF1 Antibody
Description
Introduction to RCF1 Protein and Antibody Applications
RCF1 (Respiratory supercomplex factor 1) is a mitochondrial protein critical for the assembly and stability of cytochrome bc₁-cytochrome c oxidase (COX) supercomplexes in yeast and other eukaryotes . Antibodies targeting RCF1 are essential tools for studying its structural interactions, functional domains, and regulatory roles in oxidative phosphorylation. These antibodies enable detection, localization, and biochemical characterization of RCF1 in mitochondrial membranes .
Key Domains
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Hig1 Homology Domain: Conserved N-terminal region with two transmembrane segments, critical for supercomplex bridging .
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Fungi-Specific C-Terminus: Contains a coiled-coil domain implicated in COX activity modulation .
Membrane Topology
RCF1 adopts an N out-C out orientation in the inner mitochondrial membrane, with protease-accessible termini in the intermembrane space .
Mechanistic Contributions
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Bridges cytochrome bc₁ and COX complexes via interactions with Cox3 and cytochrome c₁ .
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Maintains COX enzyme activity, particularly under stress conditions (e.g., elevated temperature) .
Functional Redundancy
RCF1 and RCF2 (Aim38) exhibit overlapping roles:
| Feature | RCF1 Contribution | RCF2 Contribution |
|---|---|---|
| Supercomplex Binding | Associates with COX and bc₁ complexes | Binds independently to supercomplexes |
| Enzyme Activity | Sustains ~70% COX activity in Δrcf1 | Partial compensation in Δrcf1 mutants |
| Stress Resilience | Required for growth at 37°C | Minor role in thermal stress adaptation |
Key Experimental Uses
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Co-Immunoprecipitation: Validated RCF1 interactions with Cox3 and AAC proteins under digitonin solubilization .
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BN-PAGE Analysis: Demonstrated RCF1 comigration with COX and bc₁ supercomplexes .
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Domain-Specific Studies: Truncation variants (ΔN, ΔC) revealed the Hig1 domain is sufficient for respiratory function .
Critical Findings Using Antibodies
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RCF1 Stability: Degradation products observed in ΔC truncations suggest C-terminus stabilizes the protein .
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Crosslinking Partners: Chemical crosslinking identified interactions with multiple inner membrane proteins, independent of RCF2 .
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Activity Modulation: Antibody-based assays showed RCF1 binding near Cox3 induces structural changes in COX’s catalytic site .
Therapeutic Potential
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Dysregulation of RCF1 is linked to COX deficiency, a hallmark of mitochondrial disorders .
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Antibodies enable screening for supercomplex assembly defects in disease models.
Unresolved Questions
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
