Recombinant Thermosipho melanesiensis Protein CrcB homolog (crcB)
Description
Introduction to Recombinant Thermosipho melanesiensis Protein CrcB Homolog
The Recombinant Thermosipho melanesiensis Protein CrcB homolog, often referred to by its gene name crcB, is a protein derived from the bacterium Thermosipho melanesiensis. This protein is part of a larger family of proteins associated with fluoride riboswitches, which are RNA structures that regulate gene expression in response to fluoride ions. The crcB gene and its protein product have been implicated in various cellular processes, including fluoride resistance and possibly membrane transport functions.
Amino Acid Sequence
The amino acid sequence of the Recombinant Thermosipho melanesiensis Protein CrcB homolog is provided as follows: MKFILIAIGGAFGALFRYFVSKVFNTHFPFNYIPLGTVIVNVLGAFLLSFVLFSSIERFE VNPNFVLFFGTGFLGAFTTFSTFAYETLSLFLTSPFRALVYFFANLFFGFFAAFFGMVLG RGKFL . This sequence is crucial for understanding the protein's structure and potential functions.
Association with Fluoride Riboswitches
Fluoride riboswitches, formerly known as the crcB RNA motif, are conserved RNA structures found in bacteria and archaea that sense fluoride ions and regulate gene expression accordingly . These riboswitches activate genes involved in fluoride resistance, including those encoding CrcB proteins. The presence of fluoride riboswitches indicates that many organisms naturally encounter toxic levels of fluoride and have evolved mechanisms to counteract its effects.
Genetic Knockout Studies
Studies involving genetic knockouts of crcB in other organisms have shown that these proteins are essential for survival in environments with high fluoride concentrations. For example, an E. coli strain lacking the crcB gene exhibited impaired growth at fluoride concentrations that were not toxic to wild-type cells . Similar findings have been reported in Pseudomonas putida, where the absence of crcB significantly increased sensitivity to sodium fluoride .
Recombinant Protein Production
The Recombinant Thermosipho melanesiensis Protein CrcB homolog is produced as a recombinant protein, available in various sizes, with a recommended storage buffer and conditions to maintain its stability . This availability facilitates further research into its structure, function, and potential applications.
Protein Characteristics
| Characteristic | Description |
|---|---|
| Species | Thermosipho melanesiensis (strain BI429 / DSM 12029) |
| Uniprot Number | A6LJF8 |
| Tag Info | Determined during production |
| Storage Buffer | Tris-based buffer, 50% glycerol |
| Storage Conditions | -20°C or -80°C for extended storage |
Amino Acid Sequence Details
| Sequence Segment | Amino Acids |
|---|---|
| Full Sequence | MKFILIAIGGAFGALFRYFVSKVFNTHFPFNYIPLGTVIVNVLGAFLLSFVLFSSIERFE VNPNFVLFFGTGFLGAFTTFSTFAYETLSLFLTSPFRALVYFFANLFFGFFAAFFGMVLG RGKFL |
Product Specs
Note: While we prioritize shipping the format currently in stock, please specify your format preference during order placement for customized preparation.
Note: Our proteins are shipped with standard blue ice packs. Dry ice shipping requires prior arrangement and incurs additional charges.
If you require a specific tag type, please inform us; we will prioritize its development.
Target Background
KEGG: tme:Tmel_0182
STRING: 391009.Tmel_0182
Q&A
FAQs for Researchers on Recombinant Thermosipho melanesiensis Protein CrcB Homolog (crcB)
Advanced Research Questions
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How to resolve discrepancies in CrcB ion selectivity across homologs?
Conflicting reports on CrcB’s substrate specificity (fluoride vs. chloride) require systematic validation:-
Electrophysiology: Perform planar lipid bilayer assays with purified protein to measure ion currents under varying pH and ion concentrations.
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Competitive binding assays: Use ICP-MS to quantify ion uptake in liposomes reconstituted with CrcB .
Example data table:
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What strategies confirm CrcB’s role in T. melanesiensis stress adaptation?
Given the organism’s hydrothermal vent habitat :-
Gene knockout: Use homologous recombination to delete crcB and compare growth under high fluoride/chloride stress vs. wild type.
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Transcriptomics: Profile gene expression changes in ΔcrcB strains under ion stress to identify compensatory pathways (e.g., upregulated efflux pumps).
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Structural dynamics: Apply cryo-EM to visualize conformational changes in CrcB at 70°C, mimicking native conditions .
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How to address low yields of functional CrcB during purification?
Common issues and solutions from homolog studies :-
Detergent screening: Test n-dodecyl-β-D-maltoside (DDM) vs. lauryl maltose neopentyl glycol (LMNG) for membrane protein stabilization.
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Buffer optimization: Include 10% glycerol and 150 mM NaCl in lysis/wash buffers to prevent aggregation.
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Validation: Confirm functionality via thermostability assays (e.g., DSC) and ligand-binding ITC.
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