Recombinant Oncorhynchus mykiss Shaker-related potassium channel tsha2
Description
Definition and Molecular Overview
Recombinant Oncorhynchus mykiss Shaker-related potassium channel tsha2 (UniProt ID: Q9I829) is a voltage-gated potassium channel protein expressed heterologously in Escherichia coli. This full-length (1-483 amino acids) protein is fused with an N-terminal polyhistidine (His) tag for purification and detection purposes . It belongs to the Shaker family of potassium channels, which are critical for regulating membrane excitability in neurons and muscle cells by mediating potassium ion efflux during action potential repolarization .
Amino Acid Sequence
The protein sequence begins with MTVVSCEIQDETVVVSPLYQDD... (full sequence available in ). Key structural features include:
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Voltage-sensing domain: Transmembrane helices (S1–S4) for detecting membrane potential changes.
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Pore-forming domain (S5–S6): Facilitates potassium ion selectivity and conductance .
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C-terminal regulatory domain: Modulates channel inactivation kinetics .
Expression and Purification
The protein is produced in E. coli as inclusion bodies, solubilized using denaturants, and refolded via gradual buffer exchange . Key steps include:
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Affinity chromatography: Ni²⁺-chelating resin for His-tag purification.
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Enterokinase cleavage: Removal of the Trx fusion partner.
Reconstitution Guidelines
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Solubility: Reconstitute in deionized water (0.1–1.0 mg/mL).
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Stability: Add 5–50% glycerol for long-term storage at -80°C .
Cardiac and Neurological Studies
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Cardiac aging: Knockdown of Shaker homologs in Drosophila increases arrhythmia susceptibility and reduces lifespan, suggesting conserved roles in cardiac stress responses .
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Mitochondrial homeostasis: Shaker channels regulate Ca²⁺ influx and ATP production, impacting cellular stress resilience .
Toxicology and Drug Development
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Channel blockers: ShK toxin (from sea anemones) selectively inhibits Kv1.3 channels, a strategy explored for autoimmune disease therapies .
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Trafficking assays: High-throughput screens (e.g., for KCNH2 variants) validate tsha2’s utility in studying channelopathies .
Comparative Analysis with Related Channels
Future Research Directions
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Functional characterization: Patch-clamp studies to define tsha2’s voltage dependence and pharmacological profile.
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Structural biology: Cryo-EM analysis to resolve pore architecture and ligand-binding sites.
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Disease modeling: Investigate tsha2’s role in piscine cardiac pathologies or environmental stress adaptations .
Product Specs
Please note: We prioritize shipping the format currently in stock. However, if you have specific format requirements, please indicate them during order placement. We will fulfill your request to the best of our ability.
Please note: Our proteins are shipped with standard blue ice packs. If you require dry ice shipping, please inform us in advance. Additional fees will apply.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. Lyophilized form has a shelf life of 12 months at -20°C/-80°C.
