Recombinant Anaeromyxobacter dehalogenans Large-conductance mechanosensitive channel (mscL)
Description
General Information
Recombinant Anaeromyxobacter dehalogenans Large-conductance mechanosensitive channel (MscL) refers to the MscL protein produced using recombinant DNA technology, derived from the bacterium Anaeromyxobacter dehalogenans . A. dehalogenans is a bacterium known for its ability to reduce metals and halo-organic compounds, found in diverse environments such as soil and sediments . MscL, or mechanosensitive channel of large conductance, is a type of protein channel that responds to mechanical stimuli, such as changes in membrane tension .
Anaeromyxobacter dehalogenans
Anaeromyxobacter dehalogenans is a bacterium belonging to the delta-Proteobacteria class and is closely related to myxobacteria . It exhibits versatile metabolic capabilities, including anaerobic respiration and the ability to reduce metals such as Fe(III) and U(VI) . Phylogenetic analysis indicates that A. dehalogenans is nested within the Myxococcales order, sharing a closer relation to Myxococcus xanthus than other myxobacteria suborders . Its genome contains genes associated with both aerobic and anaerobic lifestyles, suggesting an evolutionary adaptation from aerobic ancestors .
Large-conductance Mechanosensitive Channel (MscL)
MscL is a membrane protein that functions as a mechanosensitive channel, opening in response to mechanical stimuli such as membrane stretching . These channels are found in various organisms and play a role in protecting cells from osmotic shock by releasing solutes . The MscL from Anaeromyxobacter dehalogenans is a protein with a specific amino acid sequence, and its recombinant form is often used in research .
Recombinant Production
The recombinant form of Anaeromyxobacter dehalogenans MscL is produced using genetic engineering techniques, where the gene encoding the MscL protein is inserted into a host organism (e.g., E. coli) for expression and purification . This allows for the production of large quantities of the protein for research purposes .
Characteristics of Recombinant Anaeromyxobacter dehalogenans MscL
Research Applications
Recombinant Anaeromyxobacter dehalogenans MscL is utilized in various research applications, including:
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Structural Biology: Studying the structure and function of mechanosensitive channels .
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Biophysics: Investigating the mechanisms of channel gating and ion conductance .
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Biotechnology: Developing novel biosensors and drug delivery systems .
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Evolutionary Studies: Understanding the evolutionary relationships and adaptations of bacteria in diverse environments .
Product Specs
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Target Background
A mechanosensitive channel that opens in response to membrane lipid bilayer stretch forces. It may play a regulatory role in cellular osmotic pressure changes.
KEGG: acp:A2cp1_3746
Q&A
FAQs for Researchers on Recombinant Anaeromyxobacter dehalogenans Large-Conductance Mechanosensitive Channel (mscL)
Advanced: How do structural discrepancies in MscL oligomerization impact functional studies?
Data Contradiction Analysis:
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Homopentameric vs. alternative oligomeric states: Cryo-EM and X-ray crystallography typically show pentameric structures , but aberrant oligomerization may occur in non-native expression systems.
Basic: What are optimal expression and purification strategies for recombinant mscL?
Methodological Answer:
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Expression system: E. coli BL21(DE3) with induction at OD₆₀₀ ≈ 0.6–0.8 (16°C, 0.1 mM IPTG) .
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Purification:
Advanced: How do Anaeromyxobacter mscL homologs differ functionally from E. coli MscL?
Comparative Analysis:
Methodology: Perform cross-species electrophysiology under identical buffer conditions to directly compare activation thresholds.
Basic: How to troubleshoot low recombinant mscL solubility?
Methodological Answer:
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Codon optimization: Adjust rare codons in Anaeromyxobacter sequences for E. coli expression .
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Detergent screening: Test n-dodecyl-β-D-maltoside (DDM) or lauryl maltose neopentyl glycol (LMNG) during solubilization .
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Truncation constructs: Express cytoplasmic or transmembrane domains separately (e.g., 1-140aa vs. 1-120aa) .
Advanced: Can mscL be engineered for synthetic biology applications?
Research Design:
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Hybrid channels: Fuse mscL with optogenetic domains (e.g., light-sensitive actuators) for controlled gating .
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Biosensors: Embed mscL in lipid bilayers with fluorophore-tagged subunits to visualize real-time membrane tension changes .
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Validation: Use single-molecule FRET to confirm conformational changes upon stimulation.
Basic: How to confirm proper folding of recombinant mscL?
Methodological Answer:
