Recombinant Pan troglodytes Sodium/potassium-transporting ATPase subunit beta-1 (ATP1B1)
Description
Ion Transport Regulation
ATP1B1 ensures proper folding and plasma membrane trafficking of Na+/K+-ATPase α-subunits, maintaining electrochemical gradients essential for :
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Cellular osmoregulation
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Nerve/muscle electrophysiology
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Sodium-coupled nutrient absorption
Antiviral Innate Immunity
Recent studies demonstrate ATP1B1’s role in enhancing antiviral responses:
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Binds TRAF3/TRAF6 to amplify TBK1 and TAK1 phosphorylation, boosting IFN-β and ISG production
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Knockdown reduces IFN-stimulated genes (ISGs) by 40–60%, increasing viral replication susceptibility
Disease Associations
Experimental Uses
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Viral Pathogenesis Studies: Investigating ATP1B1-TRAF3/6 interactions in DNA/RNA virus evasion mechanisms
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Cancer Biomarker Research: Assessing ATP1B1 methylation status in RCC prognosis
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Hypertension Models: Evaluating 3’UTR variants in primate blood pressure regulation
Technical Advantages
Product Specs
Note: We will prioritize shipping the format currently in stock. However, if you have specific format requirements, please specify them when placing your order and we will accommodate your needs.
Note: All protein shipments are standardly accompanied by blue ice packs. If you require dry ice shipment, please inform us in advance as additional fees will apply.
Generally, the shelf life of the liquid form is 6 months at -20°C/-80°C. The shelf life of the lyophilized form is 12 months at -20°C/-80°C.
Target Background
Q&A
What expression systems are optimal for producing functional recombinant Pan troglodytes ATP1B1, and what purification challenges exist?
Recombinant ATP1B1 requires mammalian expression systems (e.g., HEK293 or CHO cells) due to its transmembrane domain and post-translational modifications. Prokaryotic systems like E. coli often fail to produce properly folded protein, as ATP1B1 requires β-subunit-specific glycosylation for Na+/K+-ATPase complex assembly . Key purification challenges include:
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Detergent selection: Use n-dodecyl-β-D-maltoside (DDM) to maintain transmembrane stability while avoiding protein aggregation.
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Affinity tagging: Histidine tags may interfere with native conformation; instead, employ Strep-tag II systems for improved compatibility with lipid bilayers .
A typical yield ranges from 0.5–2 mg/L culture, with purity >90% achievable via sequential immobilized metal affinity chromatography (IMAC) and size-exclusion chromatography (SEC) .
How do researchers validate the structural integrity of recombinant ATP1B1 in vitro?
A multi-modal validation pipeline is essential:
Discrepancies between predicted and observed molecular weights (e.g., 34 kDa calculated vs. 55 kDa on SDS-PAGE) typically arise from glycosylation .
What experimental strategies resolve contradictions in ATP1B1’s role across disease models?
Recent studies report conflicting roles:
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Pro-survival in alveolar epithelium: ATP1B1 stabilizes HSP90AB1 interactions, enhancing barrier function (A549 cells) .
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Oncogenic in leukemia: High expression correlates with poor prognosis (HR = 2.1, 95% CI: 1.3–3.4) .
Resolution methodology:
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Context-specific knockdown: Use CRISPR/Cas9 in isogenic cell lines to compare epithelial vs. hematopoietic systems.
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Interactome mapping: Co-IP-MS in A549 cells identified 159 ATP1B1 partners, including HSP90AB1 (fold change = 4.2) , whereas leukemia models show STAT1 co-regulation (β = 0.67, p < 0.01) .
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Pathway enrichment: Epithelial models emphasize protein folding (GO:0006457, p = 3×10⁻⁵) , while leukemic models show cell cycle dysregulation (KEGG:04110, p = 0.002) .
How to design CRISPR-edited models for studying ATP1B1’s non-canonical roles in cell adhesion?
Stepwise protocol:
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Guide RNA design: Target exon 2 (chr1:169,349,102–169,349,125, GRCh38) to disrupt β1-α subunit binding.
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Phenotypic validation:
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Rescue experiments: Co-transfect ATP1B1-CFP and α1-YFP constructs to quantify FRET efficiency (>15% indicates functional complex restoration) .
What proteomic approaches characterize ATP1B1’s interactome under pathological stress?
Advanced workflow from recent studies :
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Co-IP-MS: Anti-ATP1B1 antibody (ABclonal, A10284) with crosslinker (DSS, 2 mM).
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PRM validation: Target 6 key interactors (HSP90AB1, EIF4A1, etc.) with heavy isotope-labeled peptides.
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Network analysis:
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Module 1: Ribosomal proteins (37 nodes, score = 37)
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Module 2: Heat shock proteins (6 nodes, score = 6)
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Key finding: HSP90AB1 knockdown reduces ATP1B1 stability by 72% (p = 0.008), confirming functional dependency .
How does ATP1B1’s expression profile inform biomarker development for respiratory/cancer therapies?
| Disease | Expression Pattern | Clinical Correlation |
|---|---|---|
| ARDS | ↓50% in alveolar epithelium | Correlates with edema clearance (r = 0.81) |
| CN-AML | ↑3.2-fold vs. normal | OS HR = 1.9 (95% CI: 1.2–3.0) |
Validation strategy:
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Multiplex IHC: Co-stain ATP1B1 with CD31 (vascular leakage) or CD34 (leukemic blasts).
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Digital pathology: QuPath analysis of H-score variance (Δ >30% indicates prognostic significance) .
Why do ATP1B1 structure determinations yield conflicting topology models?
Discrepancies arise from:
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Glycosylation artifacts: PNGase F treatment reduces extracellular domain resolution by 1.8Å .
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Detergent bias: DDM preserves helix orientation but obscures residues 89–101 vs. LMNG .
Consensus approach: Hybridize cryo-EM (EMDB-3567) with molecular dynamics simulations (NAMD 3.0) to resolve extracellular loop conformations.
