KARS Antibody
Product Specs
Target Background
- Studies have shown that mutations in KARS result in a newly identified subtype of leukoencephalopathy associated with sensorineural hearing impairment. The combined effect of reduced aminoacylation and the release of LysRS from the multiple-synthetase complex (MSC) likely underlies the pathogenesis of the KARS mutations identified in this study. PMID: 28887846
- Using the TruSight One sequencing panel, we identified two novel mutations in KARS. Both mutations, previously unreported, occur in a highly conserved region of the catalytic domain and exhibit a significant impact on KARS stability. PMID: 27891585
- Caspase-8 regulates the secretion of inflammatory lysyl-tRNA synthetase in exosomes from colorectal cancer cells. PMID: 28611052
- Research indicates that KRS promotes cell-cell and cell-extracellular matrix adhesion for migration. PMID: 26891990
- Findings reveal that enzymatically active Shiga toxins trigger the dissociation of lysyl-tRNA synthetase (KRS) from the multi-aminoacyl-tRNA synthetase complex in human macrophage-like differentiated THP-1 cells, leading to its subsequent secretion. PMID: 26643967
- KRS at the plasma membrane plays new roles in metastatic migration as a signaling inducer, causing intracellular signaling for cancer dissemination. PMID: 26091349
- tRK1 forms a complex with human enolases and interacts with tRK1 and human pre-lysyl-tRNA synthetase (preKARS2). PMID: 25918939
- Structural characteristics of the KRS-LR interaction on the cell surface have been investigated. PMID: 24983501
- Lysyl-tRNA synthetase plays an essential role in HIV replication, transcriptional regulation, and cytokine-like signaling. [review] PMID: 23972532
- The role of preKARS2 in tRNA mitochondrial import has been studied. PMID: 23799079
- The KARS variant has been identified in two families affected by DFNB89-associated autosomal-recessive nonsyndromic hearing impairment. PMID: 23768514
- The C-terminal domain of HIV-1 capsid protein serves as a surrogate for human lysyl tRNA synthetase. PMID: 23208549
- A single conformational change triggered by phosphorylation leads to multiple effects driving an exclusive switch of LysRS function from translation to transcription. PMID: 23159739
- Dual roles for motif 1 residues of human lysyl-tRNA synthetase in dimerization and packaging into HIV-1 have been identified. PMID: 23095741
- Research has unveiled a unique function of KRS in the control of cell migration and its pathological implication in metastasis. PMID: 22751010
- Data supports the hypothesis that maturation of cytoplasmic KARS precursor is necessary to reveal the potent tRNA binding properties of mitochondrial KARS. PMID: 22235746
- LysRS associates with the Pol domain of GagPol. PMID: 21763493
- Results suggest that this unique geometry, which reconfigures the LysRS tetramer from alpha(2):alpha(2) to alpha(2)beta(1):beta(1)alpha(2), is designed to control both retention and mobilization of LysRS from the multi-tRNA synthetase complex. PMID: 21536907
- The interaction between helix 7 of LysRS and helix 4 of the capsid C-terminal domain of HIV-1 Gag (HIV-CA-CTD) was studied using circular dichromism spectroscopy and molecular dynamics simulation. PMID: 21093454
- These findings highlight the contribution of KARS to the emission of one of the principal signals of immunogenic cell death, CRT exposure. PMID: 20699648
- Loss-of-function lysyl-tRNA synthetase mutations have been associated with peripheral neuropathy and Charcot-Marie-Tooth disease. PMID: 20920668
- HIV-1 Gag interacts with human lysyl-tRNA synthetase during viral assembly. PMID: 12756246
- Lysyl-tRNA synthetase is packaged into human immunodeficiency virus type 1 (HIV-1) via its interaction with Gag; this enzyme facilitates the selective packaging of tRNA(3)(Lys), the primer for initiating reverse transcription, into HIV-1. PMID: 15220430
- Analysis of the interaction between HIV-1 Gag and human lysyl-tRNA synthetase has been conducted. PMID: 16702215
- HIV-1 Vpr plays an essential role in the process of packaging mitochondrial Lysyl-tRNA synthetase. PMID: 17560997
- Further characterization of the interaction between the HIV-1 capsid domain of Gag and human LysRS has been reported using truncation constructs and point mutations in the putative interaction helices. PMID: 17724017
- A 2.3-A crystal structure of a tetrameric form of human LysRS has been presented. PMID: 18272479
- mitoKARS is the first described member of a group of mitochondrial proteins whose interaction with mutant SOD1 contributes to mitochondrial dysfunction in ALS. PMID: 18715867
Q&A
What is KARS and why are antibodies against it important for research?
KARS (lysyl-tRNA synthetase) is an enzyme that catalyzes the attachment of lysine to its cognate tRNA during protein synthesis. This process occurs in both the cytoplasm and mitochondria . KARS antibodies are crucial research tools that enable investigation of this protein's expression, localization, and function in normal cellular processes and disease states. KARS1 mutations have been associated with various pathologies including autosomal recessive nonsyndromic hearing loss, congenital visual impairment, and progressive microcephaly . These connections to human disease make KARS antibodies particularly valuable for studying pathological mechanisms.
What applications are KARS antibodies validated for?
KARS antibodies have been validated for multiple research applications, with specific performance characteristics:
The validation across multiple applications ensures reliable detection of KARS in different experimental contexts .
What types of KARS antibodies are available for research?
Research-grade KARS antibodies come in several formats, each with distinct characteristics:
Selecting the appropriate antibody type depends on the specific research question, required specificity, and intended application.
How do KARS1 mutations affect antibody production and immune function?
Recent research has revealed a significant connection between KARS1 mutations and immune dysfunction. In a comprehensive study of patients with biallelic KARS1 mutations:
| Immune Abnormality | Frequency | Implications |
|---|---|---|
| Recurrent/severe infections | 9/17 patients | Suggests compromised immune defense |
| B cell lymphopenia | 3/9 patients | Reduced B cell numbers |
| Hypogammaglobulinemia | 6/15 patients | Deficiency in IgG, IgA, or IgM |
| Impaired vaccine responses | 4/7 patients | Poor antibody response to vaccination |
Functional studies demonstrated that KARS1 mutations impair B cell metabolism, specifically causing decreased mitochondrial numbers and activity . This mechanism helps explain the antibody deficiencies observed in these patients. Five patients required immunoglobulin replacement therapy to manage their condition .
What methodologies can differentiate between cytoplasmic and mitochondrial KARS pools?
Since KARS functions in both cytoplasmic and mitochondrial compartments , distinguishing between these pools requires specialized techniques:
| Methodology | Protocol Highlights | Expected Outcome |
|---|---|---|
| Subcellular Fractionation + WB | 1. Isolate cytoplasmic and mitochondrial fractions 2. Perform Western blotting with KARS antibodies 3. Include compartment-specific markers (GAPDH for cytoplasm, VDAC for mitochondria) | Differential band intensity between fractions |
| Confocal Microscopy | 1. Co-stain with KARS antibodies and mitochondrial markers 2. Use high-resolution confocal imaging 3. Analyze co-localization quantitatively | Pearson's correlation coefficient indicating degree of mitochondrial localization |
| Immunogold Electron Microscopy | 1. Use gold-conjugated secondary antibodies 2. Examine under transmission electron microscope 3. Quantify gold particle distribution | Precise ultrastructural localization of KARS |
These approaches provide complementary information about KARS distribution between cellular compartments, which may be altered in disease states.
How can KARS antibodies be used to study the role of KARS in neurodegenerative conditions?
Given the association between KARS mutations and neurological disorders , KARS antibodies offer valuable insights into pathogenic mechanisms:
| Research Approach | Methodology | Research Value |
|---|---|---|
| Expression Analysis | Compare KARS levels in affected vs. normal neural tissues using quantitative IHC or WB | Identifies altered expression in disease |
| Co-localization Studies | Combine KARS antibodies with neuronal, glial, and mitochondrial markers | Reveals changes in subcellular distribution |
| Post-translational Modifications | Use modification-specific antibodies alongside KARS antibodies | Detects disease-associated regulatory changes |
| In vitro Models | Apply KARS antibodies to neuronal cultures expressing disease-causing mutations | Elucidates cellular consequences of mutations |
These approaches can reveal how KARS dysfunction contributes to neurodegeneration, potentially through disrupted protein synthesis in neural tissues or through non-canonical functions of KARS.
What are the optimal conditions for using KARS antibodies in Western blotting?
For reliable Western blot detection of KARS:
Expected result: A specific band at approximately 74 kDa , with potential additional bands representing alternative splice variants or post-translationally modified forms.
What validation steps ensure reliable results with KARS antibodies?
Thorough validation is critical for antibody-based research:
| Validation Approach | Implementation | Significance |
|---|---|---|
| Specificity Testing | 1. Compare signal in KARS-expressing vs. KARS-knockdown samples 2. Peptide competition assays 3. Test multiple antibodies targeting different epitopes | Confirms antibody recognizes intended target |
| Cross-reactivity Assessment | Test antibody on samples from multiple species | Determines species specificity/cross-reactivity |
| Application-specific Validation | Validate separately for each application (WB, IHC, ICC-IF) | Ensures reliability across different methods |
| Reproducibility | Compare results across multiple experiments and biological replicates | Establishes consistency of findings |
These validation steps are essential for ensuring that observed signals genuinely represent KARS protein rather than experimental artifacts or cross-reactivity.
How should researchers interpret KARS antibody results in disease studies?
When using KARS antibodies to investigate disease:
| Consideration | Methodological Approach | Interpretation Guidelines |
|---|---|---|
| Mutation Effects on Epitope | Use antibodies targeting different KARS regions | Mutations may alter antibody binding sites |
| Expression vs. Function | Complement antibody studies with enzymatic activity assays | Changes in protein level may not correlate with activity |
| Tissue Variability | Include appropriate tissue-matched controls | KARS expression varies between tissues |
| Disease Heterogeneity | Analyze samples from multiple patients | Individual variability affects expression patterns |
These considerations help prevent misinterpretation of antibody-based findings in complex disease contexts.
How can KARS antibodies contribute to understanding immune dysfunction in KARS1 mutation carriers?
KARS antibodies enable detailed investigation of immune abnormalities in patients with KARS-related diseases:
These approaches can elucidate the mechanisms underlying hypogammaglobulinemia and recurrent infections observed in patients with KARS1 mutations .
Can KARS antibodies inform development of therapeutic approaches?
While current KARS antibodies serve primarily as research tools, they provide insights that could guide therapeutic development:
The development of broadly neutralizing antibody-derived therapeutics in other fields provides conceptual frameworks that might eventually be applicable to KARS-related pathologies, though significant research would be required.
What experimental designs best address contradictory findings in KARS antibody research?
When faced with contradictory results:
| Challenge | Recommended Approach | Outcome |
|---|---|---|
| Variable Antibody Performance | 1. Use multiple antibodies against different epitopes 2. Standardize protocols across laboratories 3. Establish positive and negative controls | Distinguishes antibody-specific issues from biological variations |
| Conflicting Expression Data | 1. Employ quantitative approaches (digital pathology, computer-assisted analysis) 2. Correlate with orthogonal methods (qPCR, mass spectrometry) 3. Conduct meta-analysis of published data | Resolves inconsistencies through methodological improvements |
| Discrepant Localization | 1. Use super-resolution microscopy 2. Perform fractionation with biochemical validation 3. Apply live cell imaging when possible | Provides higher resolution data to resolve conflicts |
These strategies help address contradictions that may arise from technical variations, biological heterogeneity, or methodological differences between research groups.
How might single-cell techniques enhance KARS antibody research?
Emerging single-cell technologies offer new opportunities for KARS research:
| Technology | Application with KARS Antibodies | Research Advantage |
|---|---|---|
| Single-cell Imaging Mass Cytometry | Multiplexed detection of KARS with dozens of other proteins | Reveals cell-type specific expression patterns in heterogeneous samples |
| Spatial Transcriptomics with Antibody Detection | Combined analysis of KARS protein and mRNA | Correlates protein expression with transcriptional activity |
| Microfluidic Single-cell Western Blotting | KARS detection in individual cells | Quantifies cell-to-cell variation in expression levels |
| In situ Proximity Ligation | Detecting KARS interactions at single-molecule resolution | Maps protein interaction networks in intact cells |
These approaches can reveal heterogeneity in KARS expression and function that may be masked in bulk tissue analyses, particularly important for understanding disease mechanisms.
What opportunities exist for applying computational approaches to KARS antibody research?
Computational methods enhance antibody-based research on KARS:
| Computational Approach | Implementation with KARS Antibodies | Research Value |
|---|---|---|
| Epitope Prediction | In silico analysis of antibody binding sites on KARS | Guides selection of antibodies unaffected by disease mutations |
| Machine Learning Image Analysis | Automated quantification of IHC/IF staining patterns | Enables high-throughput, unbiased analysis |
| Network Biology | Integration of KARS interaction data with systems biology | Places KARS function in broader cellular context |
| Molecular Dynamics Simulation | Predicting effects of mutations on antibody binding | Informs interpretation of experimental results |
These computational approaches complement experimental methods, enabling more comprehensive understanding of KARS biology in health and disease.
