Recombinant Schizosaccharomyces pombe Uncharacterized protein C737.05 (SPCC737.05)

What is known about the structural features of SPCC737.05?

SPCC737.05 is an uncharacterized protein from Schizosaccharomyces pombe (strain 972 / ATCC 24843) with 264 amino acids in its full-length form. The protein's amino acid sequence (MQNELNPILLSQNSIRFATRVAIFFIIRDELVEAVTWRHPVKSMCLGLTITLLYLHPVSFSAILLLVFLTMMPISMTHDVTTNLKDLQNFMASYSSSYDQLLYFRQNYYHHITPSAISSGLLLVSLVLIFLLAYLRISIDRYLPIAIWIGLISLHPKLRSYLIQFYSAKRDHVPYLQIRNELAQVWRHVDISGSQTTTRYTSFPKFNPENSVTSLDLVEPPENYSWAPQSDWTFVPPNEFRRFILWSPQPPKMNRKSSHGSNLPL) suggests multiple hydrophobic regions that could indicate transmembrane domains . When designing experiments, researchers should consider these potential membrane-spanning regions as they may influence protein folding, purification strategies, and functional assays.

How should SPCC737.05 recombinant protein be stored to maintain stability?

For optimal stability, SPCC737.05 recombinant protein should be stored in a Tris-based buffer with 50% glycerol . For extended storage, maintain the protein at -20°C or -80°C, with the latter being preferable for long-term applications. Working aliquots can be stored at 4°C for up to one week to minimize freeze-thaw cycles . It is recommended to prepare multiple small-volume aliquots upon initial thawing to prevent protein degradation, as repeated freezing and thawing should be avoided .

What are the recommended experimental controls when working with SPCC737.05?

When designing experiments with SPCC737.05, implement both positive and negative controls to ensure result validity. For functional assays, include well-characterized S. pombe proteins of similar size/structure as positive controls. For negative controls, use either buffer-only samples or proteins from non-related pathways. According to experimental design principles, variables must be clearly defined with appropriate constants maintained throughout the experiment . A proper experimental design worksheet should include:

How can researchers effectively design experiments to characterize the function of SPCC737.05?

Designing robust experiments for characterizing SPCC737.05 requires a systematic approach. Begin with a clear hypothesis about the protein's potential function based on sequence analysis and known features . Structure your experimental approach using the following framework:

• Define specific questions about SPCC737.05 function

• Identify dependent and independent variables

• Establish experimental constants

• Document safety considerations

• Develop detailed procedures with both written and visual elements

• List required materials and equipment

• Determine data collection methods and analysis strategies

For SPCC737.05, consider leveraging S. pombe cell cycle regulatory networks to identify potential functional relationships, as demonstrated in previous regulatory network studies . Analyzing time-course expression data across cell cycle phases may reveal coordination patterns with known proteins like forkhead transcription factor fkh2 .

What approaches are recommended for studying protein-protein interactions involving SPCC737.05?

To investigate protein-protein interactions of SPCC737.05, employ multiple complementary techniques:

Yeast two-hybrid screening can identify potential interaction partners, particularly when using the full-length protein or specific domains as bait. Co-immunoprecipitation experiments with tagged versions of SPCC737.05 can validate interactions in more native contexts. For higher confidence results, implement quantitative proteomic approaches using mass spectrometry.

When analyzing interaction data, consider the regulatory network context of S. pombe's cell cycle. Previous research has identified over 500 putative regulatory targets and detected many phase-specific regulatory motifs in S. pombe . Integrating your interaction data with existing regulatory networks can provide valuable context for understanding SPCC737.05's functional role.

How might SPCC737.05 be involved in S. pombe cell cycle regulation based on expression pattern analysis?

To investigate SPCC737.05's potential role in cell cycle regulation, analyze its expression patterns across different cell cycle phases using synchronized cultures. Previous research on S. pombe cell cycle has revealed that genes with similar functions often display coordinated expression patterns and can be clustered based on their temporal profiles .

For systematic analysis, implement the following approach:

• Generate synchronized S. pombe cultures using established methods

• Collect samples at regular intervals covering the entire cell cycle

• Measure SPCC737.05 expression using RT-qPCR or RNA-seq

• Apply computational methods similar to those used in previous studies that identified 31 clusters of co-regulated genes in S. pombe

• Compare SPCC737.05 expression patterns with known cell cycle regulators

What advanced statistical approaches should be applied when analyzing experimental data related to SPCC737.05?

For robust statistical analysis of SPCC737.05 experimental data, implement advanced experimental design and statistical methods as outlined in Volume 2: Advanced Experimental Design frameworks . Consider the following approaches:

What are common challenges in recombinant SPCC737.05 expression and purification, and how can they be addressed?

When working with recombinant SPCC737.05, researchers often encounter several challenges:

The amino acid sequence of SPCC737.05 reveals multiple hydrophobic regions that may form transmembrane domains , potentially leading to solubility issues during expression and purification. Consider expressing the protein in membrane-mimetic environments or using solubility-enhancing fusion tags.

How can researchers differentiate between experiment-specific artifacts and genuine characteristics of SPCC737.05?

To distinguish between artifacts and genuine characteristics:

• Implement multiple detection methods for key observations

• Include appropriate positive and negative controls in all experiments

• Perform independent biological replicates using different protein batches

• Test observations under varying experimental conditions

• Compare results from different expression systems or fusion tags

When characterizing membrane proteins like SPCC737.05 (based on its hydrophobic sequence ), artificial aggregation can be misinterpreted as oligomerization. Cross-validate findings using both detergent-solubilized and liposome-reconstituted forms of the protein to ensure observations represent native properties.

How might genomic diversity in S. pombe strains affect SPCC737.05 function and evolution?

Investigating SPCC737.05 across different S. pombe strains can provide insights into its evolutionary conservation and functional importance. A geographically diverse collection of S. pombe isolates has revealed significant genetic diversity across 40 unique haplotypes , offering an opportunity to examine SPCC737.05 sequence conservation.

The collection encompasses strains from diverse geographic locations including Mexico, Brazil (Belo Horizonte, Viçosa, Aracaju, and Salinas), as shown in the table below:

Comparative analysis of SPCC737.05 across these diverse strains could reveal:

• Conserved domains indicating functional importance

• Variable regions suggesting adaptation to different environments

• Correlations between sequence variations and phenotypic differences

• Evolutionary selection pressures on different protein domains

How can integrative multi-omics approaches advance our understanding of SPCC737.05 function?

To comprehensively characterize SPCC737.05, implement an integrative multi-omics approach that combines:

• Genomics: Analyze sequence conservation across S. pombe strains and related species

• Transcriptomics: Examine expression patterns under various conditions and across cell cycle phases

• Proteomics: Identify post-translational modifications and interaction partners

• Metabolomics: Detect metabolic changes in knockout/overexpression strains

• Phenomics: Assess cellular phenotypes when SPCC737.05 is perturbed

Previous research demonstrates the value of integrative approaches, having constructed a global gene regulatory network for S. pombe that identified phase-specific time-lagged correlations between transcription factors and genes . Similar approaches could place SPCC737.05 within this regulatory framework to elucidate its function.

When designing these multi-omics experiments, follow systematic planning principles that clearly define hypotheses, backgrounds, questions, variables, constants, and safety considerations .