Recombinant Escherichia coli O8 UPF0060 membrane protein ynfA (ynfA)
Description
Introduction to Recombinant Escherichia coli O8 UPF0060 Membrane Protein ynfA (ynfA)
The Recombinant Escherichia coli O8 UPF0060 membrane protein ynfA (ynfA) is a protein expressed in Escherichia coli and belongs to the UPF0060 family. It is a membrane protein with a full length of 108 amino acids, fused to an N-terminal His tag for easier purification and identification . This protein is often used in research settings to study membrane protein functions and interactions.
Research Findings and Applications
While specific research on the Recombinant Escherichia coli O8 UPF0060 membrane protein ynfA (ynfA) is limited, studies on similar proteins suggest that membrane proteins like ynfA play crucial roles in bacterial cell membrane functions, including transport and signaling. The His-tagged version facilitates its use in biochemical assays and structural studies.
Similar Proteins and Their Functions
Proteins similar to ynfA, such as efflux transporters, are known to contribute to bacterial resistance against antimicrobial compounds. For example, YnfA in Shigella flexneri acts as an efflux pump, aiding in the transport of cationic compounds like ethidium bromide and acriflavine, thus conferring resistance .
Expression and Purification
Recombinant Escherichia coli O8 UPF0060 membrane protein ynfA (ynfA) is typically expressed in E. coli using standard recombinant DNA technology. The His tag allows for efficient purification using nickel affinity chromatography. This method is widely used for producing high-purity membrane proteins for structural and functional studies.
Product Specs
Note: While we prioritize shipping the format currently in stock, please specify your format preference in order notes for customized preparation.
Note: All proteins are shipped with standard blue ice packs. Dry ice shipping requires prior arrangement and incurs additional charges.
The tag type is determined during the production process. If you require a specific tag, please inform us, and we will prioritize its development.
Target Background
KEGG: ecr:ECIAI1_1632
Q&A
Basic Research Questions
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What is Recombinant Escherichia coli O8 UPF0060 membrane protein ynfA (ynfA)?
Recombinant YnfA is an efflux transporter protein belonging to the Small Multidrug Resistance (SMR) family. It is characterized as an 11.9 kDa integral inner membrane protein comprised of 108 amino acids. The recombinant form refers to the protein that has been expressed using genetic engineering techniques, typically with an N-terminal His-tag to facilitate purification and downstream applications. This protein functions as an active multidrug transporter that contributes to antimicrobial resistance in bacterial species .
The commercially available recombinant YnfA protein (catalog number RFL18739EF) is expressed in E. coli expression systems and supplied as a lyophilized powder, making it suitable for various research applications focusing on membrane transport mechanisms and antimicrobial resistance .
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What structural features characterize the YnfA protein?
YnfA exhibits several key structural characteristics that define its function:
Structural Feature Description Size 108 amino acids, 11.9 kDa Transmembrane domains Four α-transmembrane helices Topology Integral inner membrane protein Conserved motifs Three conserved motif blocks essential for function Homology model template EmrE transporter (PDB ID: 3b61) Prediction tools TMHMM, TMpred, I-TASSER, AlphaFold The four α-transmembrane helical structure is a characteristic signature of members of the SMR superfamily. The predicted 3D structure of YnfA shows similarity to the EmrE transporter, with a coverage of 0.95 and a Normalized Z-score of 2.15, indicating good alignment and threading score .
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How is YnfA classified within bacterial transporter protein families?
YnfA is classified as a member of the Small Multidrug Resistance (SMR) superfamily of transporters. Phylogenetic analysis has revealed that YnfA is a distant homolog of other SMR family members and should be considered as a separate subfamily (YnfA family) alongside the three previously known subfamilies .
This classification is significant because:
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It establishes YnfA as evolutionarily distinct from other SMR transporters
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It suggests potentially unique functional properties despite shared structural features
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It indicates YnfA may have evolved specialized transport capabilities in different bacterial species
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It provides a framework for understanding related transporters in different organisms
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What are the recommended storage and handling conditions for recombinant YnfA protein?
For optimal research outcomes, recombinant YnfA protein requires specific handling protocols:
Parameter Recommendation Long-term storage -20°C/-80°C upon receipt; aliquoting necessary for multiple use Working storage 4°C for up to one week Storage buffer Tris/PBS-based buffer, 6% Trehalose, pH 8.0 Form Lyophilized powder Reconstitution Deionized sterile water to 0.1-1.0 mg/mL Cryopreservation Add 5-50% glycerol (final concentration) Important note Repeated freezing and thawing is not recommended Prior to opening, it is recommended to briefly centrifuge the vial to bring contents to the bottom. For long-term storage after reconstitution, aliquots should be prepared with glycerol added as a cryoprotectant .
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Which bacterial species contain YnfA homologs and how conserved is the protein?
YnfA is relatively widespread among Gram-negative bacteria, particularly among enterobacterial pathogens. Homology analysis reveals:
Bacterial Genus Similarity to S. flexneri YnfA Escherichia High similarity Salmonella High similarity Citrobacter High similarity Klebsiella High similarity Yersinia High similarity Multiple-sequence alignment of these homologs demonstrates that the majority of amino acids are conserved across different Gram-negative bacteria. This high degree of conservation suggests the functional importance of YnfA in these organisms. Phylogenetic analysis indicates that the YnfA protein sequence of S. flexneri closely resembles the YnfA proteins found in these related bacterial species .
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How does YnfA compare structurally and functionally to other SMR family proteins?
YnfA shares structural similarities with other SMR family proteins but also exhibits unique characteristics:
Feature YnfA Other SMR Proteins (e.g., EmrE) Size 108 amino acids, 11.9 kDa Typically 100-120 amino acids Transmembrane domains Four α-helices Four α-helices Phylogenetic classification Forms distinct YnfA subfamily Belong to established SMR subfamilies Structural homology Similar to EmrE Vary in substrate specificity Conserved motifs Three conserved blocks Share similar conserved motifs Function Efflux of specific antimicrobials Transport various substrates While YnfA maintains the core structural features of SMR transporters, phylogenetic analysis positions it as a distant homolog of other SMR family members. This suggests that YnfA may have evolved specialized functions while preserving the fundamental transport mechanism characteristic of the SMR superfamily .
