RRF1 Antibody
Description
Introduction to RRF1 Antibody
The RRF1 Antibody is a polyclonal antibody developed to detect and study Rrf1p, a mitochondrial ribosome recycling factor encoded by the RRF1 gene in Saccharomyces cerevisiae. This antibody is critical for investigating mitochondrial protein synthesis, respiratory competence, and genome stability in yeast . Rrf1p shares homology with bacterial ribosome recycling factor (RRF) and is essential for disassembling post-termination complexes in mitochondrial translation .
Antibody Production
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Antigen: A recombinant truncated Rrf1p (194 C-terminal residues) expressed in Escherichia coli with a His-tag .
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Host: Rabbit polyclonal antibody produced by Cocalico Biologicals Inc. .
Specificity Validation
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Western Blot: Detected a 23 kDa band in wild-type yeast strains and Δrrf1 strains complemented with RRF1 plasmids, but not in Δrrf1 controls .
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Mitochondrial Localization: Confirmed via subcellular fractionation and immunofluorescence, aligning with Rrf1p’s role in mitochondrial translation .
Functional Studies
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Mitochondrial Protein Synthesis: Demonstrated that Rrf1p-L209P mutation inhibits mitochondrial (but not cytoplasmic) protein synthesis by 90% at 36°C .
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Respiratory Competence:
Genomic Stability
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Mitochondrial DNA Maintenance: Δrrf1 strains exhibit rho⁻/rho⁰ (petite) phenotypes due to mitochondrial genome instability .
Critical Mutational Analysis
| Mutation | Phenotype at 36°C | Mitochondrial Protein Synthesis | Citation |
|---|---|---|---|
| L209P | Respiratory deficiency | 90% inhibition | |
| K67R/L206E | No defect | Unaffected |
Key Yeast Strains
| Strain | Genotype | Phenotype | Source |
|---|---|---|---|
| WY344 (RRF1) | MATα ino1 ura3-52 leu2-3,112 trp1::URA3 | Respiratory competent | |
| WY347 (Δrrf1) | RRF1::TRP1 | rho⁻ petite |
Plasmid Constructs
| Plasmid | Description | Function |
|---|---|---|
| pRRF1W1 | Low-copy RRF1 complementation | Restores respiration |
| prrf1-L209P | Temperature-sensitive rrf1 allele | Conditional knockout |
Cross-Reactivity and Limitations
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Specificity: No cross-reactivity with cytoplasmic or chloroplast proteins confirmed .
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Limitations: Weak signals in wild-type strains due to low endogenous Rrf1p levels .
Significance in Mitochondrial Biology
The RRF1 Antibody has enabled critical discoveries, including:
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Evolutionary Insights: Conservation of ribosome recycling mechanisms between mitochondria and bacteria .
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Therapeutic Potential: Structural differences between Rrf1p and bacterial RRF suggest targets for antibacterial agents .
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Disease Modeling: Links between mitochondrial translation defects and human metabolic disorders .
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
KEGG: ago:AGOS_AAL074C
STRING: 33169.AAS50292
