Recombinant Archaeoglobus fulgidus Uncharacterized protein AF_1578 (AF_1578)

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Description

Amino Acid Sequence

The full-length protein spans residues 1–77, with the sequence:
MIFLIISVPFGIYSLVIYNLTRRAPGKMRYLIPPLLTATLPALYLPLTGFKVSYDLLPVVGYLTYSQFLLLLLQLQR .

PropertySpecification
Source OrganismArchaeoglobus fulgidus (hyperthermophilic sulfate-reducing archaeon)
Expression HostE. coli
TagN-terminal His-tag
FormLyophilized powder
Purity>90% (SDS-PAGE)
Storage BufferTris/PBS-based buffer, 6% trehalose, pH 8.0

Key Features

  • Thermostability: Reflects the organism’s optimal growth at 83°C .

  • Purification: His-tag enables affinity chromatography for high-yield isolation .

Production and Handling

Genomic and Metabolic Insights

  • Host Organism: A. fulgidus is a sulfate-reducing archaeon with a 2.3 Mbp genome, including genes for sulfate metabolism and CRISPR elements .

  • Protein Role: AF_1578’s function remains unknown, but its presence in A. fulgidus suggests potential involvement in:

    • Thermophilic Adaptation: Structural stability under extreme heat.

    • Metabolic Pathways: Hypothetical roles in energy production or stress response, given the organism’s chemolithoautotrophic lifestyle .

Comparative Studies

While unrelated to AF_1578, A. fulgidus has been studied for DNA repair mechanisms, such as uracil-DNA glycosylase (UDG) activity in base excision repair (BER) . These studies highlight the organism’s unique biochemical adaptations, though AF_1578’s specific role in such pathways is unexplored.

Availability and Research Tools

  • Commercial Sources: Offered as lyophilized powder (e.g., Creative BioMart, CUSABIO) .

  • ELISA Kits: Available for antigen detection in research settings .

Research Gaps and Future Directions

  • Functional Characterization: No direct evidence links AF_1578 to enzymatic activity, regulatory processes, or structural roles.

  • Interactome Studies: No reported interactions with other proteins or metabolites .

  • Hypothesis Testing: Potential targets include heat shock response, redox regulation, or sulfate metabolism, informed by A. fulgidus’ genomic inventory .

Product Specs

Form
Lyophilized powder
Note: We will prioritize shipping the format currently in stock. However, if you have specific format requirements, please indicate them when placing your order, and we will fulfill your request.
Lead Time
Delivery time may vary depending on the purchase method and location. Please consult your local distributors for specific delivery times.
Note: All proteins are shipped with standard blue ice packs by default. If you require dry ice shipment, please inform us in advance, as additional fees may apply.
Notes
Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.
Reconstitution
We recommend briefly centrifuging the vial prior to opening to ensure the contents settle at the bottom. Reconstitute the protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL. For long-term storage, we suggest adding 5-50% glycerol (final concentration) and aliquoting at -20°C/-80°C. Our default final glycerol concentration is 50%. Customers can use this as a reference.
Shelf Life
Shelf life is influenced by various factors, including storage conditions, buffer composition, temperature, and the inherent stability of the protein.
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.
Storage Condition
Store at -20°C/-80°C upon receipt. Aliquoting is necessary for multiple uses. Avoid repeated freeze-thaw cycles.
Tag Info
Tag type is determined during the manufacturing process.
The tag type will be determined during the production process. If you have specific tag type requirements, please inform us, and we will prioritize developing the specified tag.
Synonyms
AF_1578; Uncharacterized protein AF_1578
Buffer Before Lyophilization
Tris/PBS-based buffer, 6% Trehalose.
Datasheet
Please contact us to get it.
Expression Region
1-77
Protein Length
full length protein
Species
Archaeoglobus fulgidus (strain ATCC 49558 / VC-16 / DSM 4304 / JCM 9628 / NBRC 100126)
Target Names
AF_1578
Target Protein Sequence
MIFLIISVPFGIYSLVIYNLTRRAPGKMRYLIPPLLTATLPALYLPLTGFKVSYDLLPVV GYLTYSQFLLLLLQLQR
Uniprot No.

Target Background

Database Links

KEGG: afu:AF_1578

STRING: 224325.AF1578

Subcellular Location
Cell membrane; Multi-pass membrane protein.

Q&A

What is Archaeoglobus fulgidus and why is AF_1578 significant for research?

Archaeoglobus fulgidus is a hyperthermophilic, sulfate-reducing archaeon commonly found in high-temperature and high-pressure marine environments, including deep-sea hydrothermal vents at depths of 2-5 km below sea level (20-50 MPa pressures) . It was the first hyperthermophilic sulfate-reducing archaeon to be isolated and characterized, and one of the first archaea to have its genome sequenced . AF_1578 is an uncharacterized protein from this organism, making it potentially valuable for understanding archaeal protein function in extreme environments.

What growth conditions are optimal for Archaeoglobus fulgidus when expressing native proteins?

Archaeoglobus fulgidus exhibits growth capabilities under various conditions. For heterotrophic metabolism (lactate oxidation coupled to sulfate reduction), it can grow at pressures up to 60 MPa, with maximum growth rates observed at 20 MPa, suggesting it is a moderate piezophile under these conditions . For autotrophic metabolism (CO₂ fixation coupled to thiosulfate reduction via H₂), it shows piezotolerance with growth rates remaining nearly constant from 0.3 to 40 MPa . The optimal temperature for growth is 83°C, making it a true hyperthermophile . These conditions are important to consider when studying native protein expression and function.

What expression systems are suitable for recombinant production of AF_1578?

Based on available information, E. coli has been successfully used as an expression host for recombinant AF_1578 protein production . The protein has been produced with a His-tag, which facilitates purification through affinity chromatography techniques . When expressing archaeal proteins in bacterial systems, researchers should consider potential issues with protein folding, post-translational modifications, and stability at mesophilic temperatures. Alternative expression systems such as archaeal hosts might be considered for proteins that prove difficult to express functionally in bacterial systems.

What is the molecular weight and basic structure of AF_1578?

AF_1578 is a relatively small protein with a length of 77 amino acids in its full-length form . While detailed structural information is not provided in the search results, its small size suggests it might function as part of a larger complex or have a specialized regulatory role. Computational prediction of secondary and tertiary structure would be valuable first steps in characterizing this protein.

How might high-pressure adaptation in Archaeoglobus fulgidus affect the structure and function of proteins like AF_1578?

Archaeoglobus fulgidus demonstrates remarkable piezotolerance and even piezophilic characteristics in some metabolic modes . For proteins from such organisms, pressure adaptation often involves structural modifications that maintain protein function under compression. These adaptations may include:

  • Increased hydrophobic core packing

  • Reduced void volumes within the protein structure

  • Strengthened ionic interactions

  • Modified amino acid composition favoring residues less affected by pressure

Experimental approaches to study pressure effects on AF_1578 should include:

  • High-pressure circular dichroism to assess secondary structure stability

  • Pressure-resolved fluorescence spectroscopy to monitor tertiary structure changes

  • Activity assays performed under pressure to correlate structural changes with function

  • Comparative analysis with homologous proteins from non-piezophilic organisms

What bioinformatic approaches would be most effective for predicting the function of AF_1578?

For uncharacterized proteins like AF_1578, a multi-faceted bioinformatic approach is recommended:

  • Sequence homology searches against characterized proteins using sensitive methods like PSI-BLAST and HMM-based tools

  • Structural prediction using AlphaFold2 or similar tools, followed by structural homology searches

  • Genomic context analysis examining neighboring genes and potential operonic organization

  • Phylogenetic profiling to identify co-occurrence patterns with other proteins across species

  • Analysis of conserved domains and motifs that might suggest biochemical function

  • Computational prediction of protein-protein interaction partners

This comprehensive approach can generate testable hypotheses about AF_1578 function that can guide experimental design.

What methods are recommended for studying potential involvement of AF_1578 in heat shock response?

Given that Archaeoglobus fulgidus has a well-characterized heat shock response involving approximately 350 genes (14% of its genome) , investigating whether AF_1578 plays a role in this response would be valuable. A methodological approach should include:

  • Transcriptomic analysis: Compare AF_1578 expression levels before and after heat shock at different time points (similar to the study described for AF1298)

  • Promoter analysis: Examine the upstream region of AF_1578 for potential heat shock regulatory elements similar to the CTAAC-N5-GTTAG motif identified for HSR1-regulated genes

  • Protein-DNA interaction assays: Use electrophoretic mobility shift assays (EMSA) and DNase I footprinting to test if known heat shock regulators like HSR1 bind to the AF_1578 promoter region

  • Gene knockout or knockdown: Assess how loss of AF_1578 affects the organism's ability to survive heat shock

  • Interactome studies: Identify proteins that physically interact with AF_1578 under normal and heat shock conditions

What experimental approaches would be suitable for determining if AF_1578 has DNA-binding capabilities similar to HSR1?

The search results describe HSR1 (AF1298) as a heat shock regulator with DNA-binding capabilities . To investigate whether AF_1578 might have similar properties:

  • Sequence and structural analysis: Look for DNA-binding motifs such as helix-turn-helix domains similar to those found in HSR1

  • Electrophoretic mobility shift assays (EMSA): Test purified recombinant AF_1578 for binding to various DNA fragments, starting with its own promoter region (to test for autoregulation) and promoters of heat shock genes

  • DNase I footprinting: If binding is observed, determine the specific DNA sequence recognized by AF_1578

  • Chromatin immunoprecipitation: In vivo identification of genomic binding sites

  • Mutational analysis: Create point mutations in potential DNA-binding regions to confirm their importance

How can researchers address the challenges of studying protein-protein interactions involving AF_1578 under extreme conditions?

Studying protein-protein interactions (PPIs) for proteins from extremophiles presents unique challenges due to the extreme conditions required for native folding and function. For AF_1578, consider these approaches:

  • High-temperature pull-down assays: Adapt conventional pull-down methods to function at elevated temperatures

  • Chemical cross-linking mass spectrometry (XL-MS): Perform in vivo crosslinking at high temperatures before cell lysis to capture native interactions

  • Yeast two-hybrid adaptations: Use thermostable variants of the system or shuttle to archaeal two-hybrid systems

  • Surface plasmon resonance (SPR) under extreme conditions: Modified instruments that can operate at high temperatures

  • Hydrogen-deuterium exchange mass spectrometry (HDX-MS) to identify interaction interfaces

  • Reconstituted systems with purified components tested under various temperature and pressure conditions

These methods should be coupled with bioinformatic predictions of potential interaction partners based on genomic context and co-expression data.

What purification strategies are optimal for obtaining high-quality recombinant AF_1578 protein?

For the purification of recombinant His-tagged AF_1578 expressed in E. coli , the following methodological approach is recommended:

  • Initial capture: Immobilized metal affinity chromatography (IMAC) using Ni-NTA or similar resin

  • Heat treatment: Exploiting the thermostability of archaeal proteins to remove mesophilic host contaminants by heating the lysate to 60-70°C

  • Secondary purification: Size exclusion chromatography to remove aggregates and ensure homogeneity

  • Quality control: Assess purity by SDS-PAGE and protein identity by mass spectrometry

  • Functional verification: Develop activity assays based on bioinformatic predictions of function

For researchers requiring protein for structural studies, additional considerations include buffer optimization to enhance stability and reduce aggregation.

How should experiments be designed to study the effects of temperature and pressure on AF_1578 stability and function?

Given that Archaeoglobus fulgidus is both thermophilic and piezotolerant/piezophilic , experiments studying AF_1578 should account for these environmental factors:

  • Temperature-dependent studies:

    • Thermal stability assays using differential scanning calorimetry or thermofluor assays

    • Activity measurements across a temperature range (60-90°C)

    • Structural characterization at different temperatures using circular dichroism or FTIR

  • Pressure-dependent studies:

    • High-pressure cultivation systems similar to those described for A. fulgidus growth studies

    • Custom pressure vessels for biochemical assays

    • Pressure-resolved spectroscopic techniques to monitor conformational changes

  • Combined temperature-pressure experiments:

    • Establish a two-dimensional stability/activity map across relevant temperature and pressure ranges

    • Design specialized equipment that can maintain both high temperature and high pressure simultaneously

What approaches can be used to identify potential physiological substrates or binding partners of AF_1578?

Since AF_1578 is uncharacterized, identifying its substrates or binding partners will be crucial for understanding its function. Consider these methodological approaches:

  • Metabolite profiling: Compare metabolomes of wild-type and AF_1578 knockout strains

  • Affinity purification: Use immobilized AF_1578 to capture interacting proteins or small molecules from cell lysates

  • Thermal proteome profiling: Identify proteins whose thermal stability changes upon binding to AF_1578

  • Library screening: Test interaction with libraries of metabolites, peptides, or nucleic acids

  • In silico docking: Computational prediction of potential binding partners based on structural models

  • Co-localization studies: If antibodies are available, use immunofluorescence to identify cellular localization that might suggest function

How can researchers overcome the challenge of expressing archaeal proteins in heterologous systems?

Expression of archaeal proteins in mesophilic hosts like E. coli often presents challenges due to differences in genetic code usage, folding environments, and post-translational modifications. To overcome these issues:

  • Codon optimization: Adjust codon usage to match the expression host

  • Fusion partners: Use solubility-enhancing fusion tags such as SUMO, MBP, or TrxA

  • Co-expression with chaperones: Include molecular chaperones from the native organism or thermostable chaperones

  • Expression conditions: Lower temperatures, slower induction, and specialized media formulations

  • Alternative hosts: Consider archaeal expression systems for particularly challenging proteins

  • Cell-free systems: Use thermostable cell-free expression systems derived from thermophiles

For AF_1578 specifically, expression in E. coli with a His-tag has been successful , suggesting that basic optimization strategies may be sufficient.

What analytical techniques are most appropriate for characterizing the structure and oligomeric state of small archaeal proteins like AF_1578?

For a small protein like AF_1578 (77 amino acids) , appropriate structural characterization techniques include:

  • X-ray crystallography: If the protein can be crystallized, this offers high-resolution structural information

  • NMR spectroscopy: Particularly suitable for small proteins, providing both structural and dynamic information

  • Cryo-electron microscopy: Especially useful if AF_1578 forms larger complexes

  • Small-angle X-ray scattering (SAXS): For low-resolution shape determination in solution

  • Analytical ultracentrifugation: To determine oligomeric state and homogeneity

  • Native mass spectrometry: For accurate mass determination of intact complexes

  • Hydrogen-deuterium exchange mass spectrometry: To probe solvent accessibility and conformational dynamics

The small size of AF_1578 makes it an excellent candidate for NMR studies, which could provide detailed structural information even in the absence of crystallization.

How might systems biology approaches contribute to understanding the role of AF_1578 in the context of Archaeoglobus fulgidus adaptation to extreme environments?

Systems biology approaches can provide a holistic understanding of AF_1578's role:

  • Multi-omics integration: Combine transcriptomics, proteomics, and metabolomics data under various stress conditions

  • Network analysis: Place AF_1578 in the context of global protein-protein interaction networks

  • Comparative genomics: Analyze the conservation and evolution of AF_1578 across related extremophiles

  • Flux balance analysis: Model the metabolic impact of AF_1578 perturbation

  • Adaptive laboratory evolution: Select for strains with altered expression of AF_1578 and characterize adaptive mutations

This integrated approach could reveal emergent properties not apparent from isolated studies and place AF_1578 in the broader context of archaeal adaptation to extreme environments.

What potential biotechnological applications might arise from characterization of AF_1578?

While avoiding commercial aspects as requested, the scientific exploration of potential applications includes:

  • Thermostable enzymes for research applications: If AF_1578 has enzymatic activity, its thermostability could be advantageous for high-temperature reactions

  • Structural biology insights: Understanding how small proteins from extremophiles maintain stability could inform protein engineering efforts

  • Biomarkers for environmental monitoring: If AF_1578 is specific to Archaeoglobus fulgidus, it could serve as a biomarker for detecting these organisms in environmental samples

  • Model systems for astrobiology: Extremophile proteins serve as models for potential extraterrestrial life

These applications represent scientific research directions rather than commercial developments.

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