Recombinant Pongo abelii UPF0420 protein C16orf58 homolog
Description
Introduction to Recombinant Pongo abelii UPF0420 Protein C16orf58 Homolog
The Recombinant Pongo abelii UPF0420 protein C16orf58 homolog is a recombinant protein derived from the Sumatran orangutan (Pongo abelii). This protein is homologous to the human C16orf58 protein, suggesting a conserved function across species. The recombinant form of this protein is produced using biotechnological methods, typically involving the insertion of the gene encoding the protein into a suitable expression vector, followed by expression in a host organism such as bacteria or mammalian cells.
Key Features:
-
Species: Pongo abelii (Sumatran orangutan)
-
Protein Type: Recombinant
-
Homolog: C16orf58
-
Quantity: Available in quantities such as 50 µg, with options for other quantities upon inquiry
-
Storage Buffer: Tris-based buffer with 50% glycerol
-
Storage Conditions: Store at -20°C for short-term storage or -80°C for long-term storage
Protein Structure and Sequence
The Recombinant Pongo abelii UPF0420 protein C16orf58 homolog has a specific amino acid sequence that defines its structure and function. The sequence provided includes a detailed arrangement of amino acids, which is crucial for understanding its biological role and interactions.
Amino Acid Sequence:
The sequence begins with MADDAGLETSLCSEQFGSGEARGCRVAADGSLQWEVGGWRWWGLSRAFTVKPEGRDSGEV GAPGAPSPPLSGLQAVFLPQGFPDSVSPDYLPYQLWDSVQAFASGLSGSLATQAVLLGIG VGNAKATVSAATATWLVKDSTGmLGRIVFAWWKGSKLDCNAKQWRLFADILNDVAMFLEI MAPVYPICFTMTVSTSNLAKCIVSVAGGATRAALTVHQARRNNMADVSAKDSSQETLVNL VGLLVSLLmLPLVSGCPGFSLGCFFFLTALHIYANYRAVRALVMETLNEGRLRLVLKHYL QRGEVLNPTAANRMEPLWTGFWPAPSLSLGVPLHRLVSSVFELQQLVEGHQEPYLLCWDQ SRNQVQVVLNQKAGPKTILRAATHGLmLGALQGDGPLPAELEELRNRVQAGPKKESWVIV KETHEVLDmLFPKFLKGLQDAGWKTEKHQLEVDEWRATWLLSPEKKVL.
Biological Function
While specific biological functions of the Recombinant Pongo abelii UPF0420 protein C16orf58 homolog are not extensively documented, its homology to human proteins suggests potential roles in cellular processes. The conservation of such proteins across species often indicates essential functions, such as involvement in cellular signaling pathways or structural roles within cells.
Potential Functions:
-
Cellular Signaling: May participate in signaling pathways that regulate cell growth, differentiation, or survival.
-
Structural Roles: Could contribute to maintaining cellular architecture or facilitating interactions between different cellular components.
Research Applications
This recombinant protein is useful in various research applications, including studies on protein function, cellular biology, and comparative genomics. It can be used in assays to understand protein-protein interactions, enzymatic activity, or as a tool for studying evolutionary conservation of protein functions.
Applications:
-
Protein-Protein Interaction Studies: Useful for identifying binding partners and understanding how these interactions affect cellular processes.
-
Enzymatic Activity Assays: If the protein has enzymatic activity, it can be used to study substrate specificity and catalytic mechanisms.
-
Comparative Genomics: Helps in understanding how protein functions evolve across different species.
Working Aliquots:
-
Store working aliquots at 4°C for up to one week.
Future Directions:
-
Functional Studies: Investigating the protein's role in cellular processes and its interactions with other proteins.
-
Comparative Studies: Comparing its function across different species to understand evolutionary conservation.
Data Table: Key Features of Recombinant Pongo abelii UPF0420 Protein C16orf58 Homolog
| Feature | Description |
|---|---|
| Species | Pongo abelii (Sumatran orangutan) |
| Protein Type | Recombinant |
| Homolog | C16orf58 |
| Quantity | 50 µg (other quantities available upon request) |
| Storage Buffer | Tris-based buffer with 50% glycerol |
| Storage Conditions | -20°C for short-term, -80°C for long-term |
| Amino Acid Sequence | MADDAGLETSLCSEQFGSGEARGCRVAADGSLQWEVGGWRWWGLSRAFTVKPEGRDSGEV GAPGAPSPPLSGLQAVFLPQGFPDSVSPDYLPYQLWDSVQAFASGLSGSLATQAVLLGIG VGNAKATVSAATATWLVKDSTGmLGRIVFAWWKGSKLDCNAKQWRLFADILNDVAMFLEI MAPVYPICFTMTVSTSNLAKCIVSVAGGATRAALTVHQARRNNMADVSAKDSSQETLVNL VGLLVSLLmLPLVSGCPGFSLGCFFFLTALHIYANYRAVRALVMETLNEGRLRLVLKHYL QRGEVLNPTAANRMEPLWTGFWPAPSLSLGVPLHRLVSSVFELQQLVEGHQEPYLLCWDQ SRNQVQVVLNQKAGPKTILRAATHGLmLGALQGDGPLPAELEELRNRVQAGPKKESWVIV KETHEVLDmLFPKFLKGLQDAGWKTEKHQLEVDEWRATWLLSPEKKVL |
References:
Product Specs
Note: While we prioritize shipping the format currently in stock, please specify your format preference during ordering for customized preparation.
Note: Our proteins are shipped with standard blue ice packs. Dry ice shipping requires prior arrangement and incurs additional charges.
The tag type is determined during production. If you require a specific tag, please inform us, and we will prioritize its development.
Target Background
KEGG: pon:100173090
UniGene: Pab.18909
Q&A
What are the structural and biochemical characteristics of Recombinant Pongo abelii UPF0420 Protein C16orf58 Homolog?
The protein is a 468-amino-acid full-length construct expressed in E. coli with an N-terminal His-tag for purification . Its molecular weight can be estimated at approximately 52–55 kDa based on sequence length, though post-translational modifications in eukaryotic systems may alter this. Researchers should validate purity and stability using SDS-PAGE, size-exclusion chromatography, and mass spectrometry.
Key parameters for experimental validation:
| Parameter | Specification |
|---|---|
| Expression System | Escherichia coli (prokaryotic host) |
| Tag | Polyhistidine (His-tag) |
| Protein Length | 1–468 residues |
| Purification Method | Immobilized metal affinity chromatography (IMAC) |
How should researchers design initial functional assays for this protein?
A three-phase experimental framework is recommended :
-
Hypothesis formulation: Define the protein’s putative role (e.g., enzymatic activity, structural binding) based on homology to human C16orf58.
-
Variable selection:
-
Independent variable: Protein concentration or incubation time.
-
Dependent variable: Measurable output (e.g., substrate conversion rate, binding affinity).
-
-
Control groups: Include tag-cleaved protein samples and empty vector lysates to account for host-cell contaminants.
For kinetic studies, use Michaelis-Menten assays with varying substrate concentrations. Normalize activity measurements against negative controls to isolate target effects .
What bioinformatics tools are essential for preliminary characterization?
Leverage homology modeling using SWISS-MODEL or AlphaFold2 to predict tertiary structure. Cross-reference conserved domains with Pfam or InterPro databases. For pathway analysis, use STRING-db to map potential interactors, though experimental validation is required due to limited annotated data for Pongo abelii proteins .
How can conflicting data on protein-protein interactions be resolved?
Discrepancies in interaction studies (e.g., yeast two-hybrid vs. co-IP results) often arise from methodological differences. Implement a tiered validation protocol:
-
Primary screening: High-throughput yeast two-hybrid or affinity pull-down assays.
-
Secondary confirmation: Co-immunoprecipitation (co-IP) under physiological buffer conditions.
-
Tertiary validation: Biolayer interferometry or surface plasmon resonance (SPR) for binding kinetics.
Case example: If pull-down assays detect interactions absent in co-IP, check for tag accessibility issues or denaturation during purification. Include controls with scrambled peptide tags to rule out nonspecific binding .
What strategies optimize crystallization trials for structural studies?
Crystallization challenges are common with poorly soluble proteins. Apply these steps:
-
Buffer optimization: Screen 20–30 conditions varying pH (5.5–8.5), ionic strength (50–500 mM NaCl), and additives (e.g., 5% glycerol).
-
Thermal shift assays: Identify stabilizers (e.g., ligands, ions) that increase melting temperature (Tm) by ≥5°C.
-
Membrane protein adaptations: If transmembrane domains are predicted, use lipid cubic phase (LCP) crystallization.
Recent advances in cryo-EM may bypass crystallization hurdles, enabling single-particle analysis at 2–3 Å resolution .
How do researchers address contradictory functional annotations across homologs?
Human C16orf58 homologs are implicated in RNA metabolism and mitochondrial function. To resolve conflicts:
-
Comparative phylogenetics: Construct a maximum-likelihood tree with OrthoFinder to identify clade-specific functional divergence.
-
Functional complementation: Knock out the ortholog in a model organism (e.g., S. cerevisiae) and test rescue efficiency with the Pongo abelii protein.
-
Multi-omics integration: Correlate transcriptomic/proteomic profiles under knockdown conditions using weighted gene co-expression network analysis (WGCNA).
Data reconciliation table:
| Discrepancy Source | Resolution Strategy |
|---|---|
| Variant enzymatic activity | Steady-state kinetics across pH 4–9 |
| Subcellular localization | Fractionation + immunofluorescence |
| Pathway enrichment conflicts | CRISPRi perturbation + RNA-seq |
What statistical models are appropriate for dose-response experiments?
Use nonlinear regression with four-parameter logistic (4PL) models:
Where = log(concentration), = response. Validate model fit via Akaike information criterion (AIC) and residual plots .
How to mitigate batch effects in large-scale production?
While commercial production is excluded from discussion, academic-scale batches require:
-
Randomized block designs: Allocate protein preps from different purification batches across experimental groups.
-
Spike-in controls: Add a fluorescently labeled protein aliquot to quantify technical variability.
-
Mixed-effects modeling: Include batch as a random effect in lme4 or BRMS packages for R.
