Recombinant Human Uncharacterized membrane protein C1orf95 (C1orf95)

What is C1orf95 and what are its known alternative names?

C1orf95, formally known as Chromosome 1 open reading frame 95, is an uncharacterized membrane protein localized to the cell membrane. The protein has several recognized synonyms including STUM (STUM_HUMAN), Hypothetical protein LOC375057, Spec3 like, and CA095_HUMAN. The gene is located on Chromosome 1, which spans approximately 260 million base pairs and constitutes about 8% of the human genome .

What are the key molecular identifiers for C1orf95?

C1orf95 can be identified using the following reference information:

• UniProt Primary Accession: Q69YW2

• UniProt Entry Name: STUM_HUMAN

• Gene Symbol: C1ORF95

• KEGG Identifier: hsa:375057

This information is essential for database searches and accurate identification across different research platforms.

What detection methods are currently available for C1orf95 research?

Several validated detection methods exist for C1orf95 research:

What sample types are suitable for C1orf95 analysis?

C1orf95 can be studied in various biological materials including tissue homogenates, cell lysates, and other biological fluids . When designing experiments, it's important to note that commercial detection kits are optimized for native protein samples rather than recombinant proteins, which may affect experimental outcomes when working with artificial expression systems .

How should researchers approach differential expression analysis of C1orf95?

When analyzing C1orf95 expression changes, researchers should implement a multi-platform validation approach:

• Initial screening can be conducted using microarray analysis, which has successfully identified C1orf95 as a differentially expressed gene (DEG) with a fold change of 2.2 in specific tissues .

• Validation should be performed using qPCR, which typically shows greater sensitivity than microarray platforms. Previous research has demonstrated that qPCR validation can confirm expression trends identified in microarray data, with approximately 71% concordance between platforms .

• Statistical analysis should account for platform differences - qPCR typically yields higher fold-change values for the same samples compared to microarray analysis due to its enhanced sensitivity .

• For comprehensive expression profiling, consider integrating data from multiple tissues or conditions to identify context-specific regulation patterns.

What experimental approaches are recommended for investigating C1orf95 function?

Given the uncharacterized nature of C1orf95, a systematic multi-method approach is recommended:

• Loss-of-function studies:

  • CRISPR/Cas9-mediated knockout in relevant cell lines

  • siRNA/shRNA knockdown with multiple targeting sequences

  • Analysis of resulting phenotypes across multiple cellular processes

• Protein interaction studies:

  • Proximity labeling techniques (BioID, APEX)

  • Co-immunoprecipitation followed by mass spectrometry

  • Membrane yeast two-hybrid assays specifically designed for membrane proteins

• Subcellular localization:

  • High-resolution microscopy with tagged constructs

  • Subcellular fractionation with immunoblotting

  • Correlation with known membrane compartment markers

• Pathway analysis:

  • Investigation of calcium signaling components, as altered intracellular ion homeostasis, particularly Ca²⁺, may be relevant based on pathways where C1orf95 shows differential expression

  • Examination of potential connections to Ras signaling or HIF-1 signaling pathways

What are the critical considerations for generating and validating recombinant C1orf95?

When producing recombinant C1orf95, researchers should consider:

• Expression system selection:

  • Mammalian systems (HEK293, CHO) may better preserve native conformation and post-translational modifications

  • Insect cell systems offer a balance between yield and proper folding

  • Bacterial systems may require optimization for membrane protein expression

• Construct design:

  • Include purification tags (His, FLAG) positioned to minimize interference with protein function

  • Consider fusion partners (MBP, SUMO) to enhance solubility

  • Include protease cleavage sites for tag removal

• Validation approaches:

  • Confirm proper membrane integration using membrane fractionation

  • Verify structural integrity using circular dichroism or limited proteolysis

  • Assess functionality through binding assays if binding partners are identified

• Potential challenges:

  • Commercial detection kits are optimized for native rather than recombinant forms of C1orf95

  • Membrane proteins often require specialized detergents for extraction and purification

  • Yield and stability may vary significantly between expression systems

How can researchers effectively design C1orf95 antibodies for specific applications?

Developing effective antibodies against C1orf95 requires careful epitope selection and validation:

• Epitope selection strategies:

  • Target unique, accessible regions based on predicted membrane topology

  • Avoid highly conserved domains that might cross-react with related proteins

  • Consider multiple epitopes to generate complementary antibodies

• Production approaches:

  • Monoclonal antibodies offer consistency between batches and high specificity

  • Polyclonal antibodies provide broader epitope recognition

  • Recombinant antibodies allow for engineered specificity and consistent production

• Validation requirements:

  • Western blotting against both recombinant and endogenous protein

  • Immunoprecipitation followed by mass spectrometry confirmation

  • Immunostaining with siRNA/CRISPR knockdown controls

  • Cross-validation between antibodies targeting different epitopes

• Application-specific considerations:

  • For ELISA: optimize coating conditions and blocking reagents

  • For immunohistochemistry: validate fixation and antigen retrieval methods

  • For immunofluorescence: confirm specificity in both fixed and live-cell applications

What bioinformatic approaches are valuable for studying C1orf95?

Several computational methods can provide insights into this uncharacterized protein:

• Sequence-based analysis:

  • Multiple sequence alignment with orthologs to identify conserved regions

  • Transmembrane topology prediction using algorithms like TMHMM or Phobius

  • Motif identification for potential functional domains

  • Signal peptide prediction for trafficking information

• Structural modeling:

  • Ab initio structure prediction using AlphaFold or RoseTTAFold

  • Molecular dynamics simulations to assess stability and flexibility

  • Binding site prediction for potential ligand interactions

• Expression correlation analysis:

  • Integration with transcriptomic datasets to identify co-expressed genes

  • Network analysis to predict functional associations

  • Tissue-specific expression pattern analysis

• Morphometric similarity mapping:

  • In neurological research contexts, morphometric similarity mapping can be valuable for correlating C1orf95 expression with structural brain patterns

  • This approach quantifies similarity between cortical areas in terms of multiple MRI parameters and has been used successfully in studies of psychosis and other neurological conditions

How can researchers investigate potential roles of C1orf95 in disease contexts?

Based on current research contexts where C1orf95 has been identified:

• In muscle-related pathologies:

  • C1orf95 showed differential expression (FC = 2.2) in studies of white striping abnormality in muscle tissue

  • Research design should incorporate physiological stress conditions that mimic pathological states

  • Consider calcium homeostasis dysregulation as a potential mechanistic link

• In neurological/psychiatric contexts:

  • Given that Chromosome 1 contains the DISC1 gene linked to schizophrenia , and morphometric similarity mapping has been used in psychosis research , investigation of C1orf95 in neuropsychiatric contexts may be valuable

  • Consider both structural and functional neuroimaging correlations

  • Implement patient-derived cellular models (iPSCs) for disease-relevant phenotyping

• Experimental design considerations:

  • Include appropriate disease and control samples with sufficient statistical power

  • Employ both in vitro and in vivo models when available

  • Utilize multi-omics approaches (transcriptomics, proteomics, metabolomics) for comprehensive characterization

  • Consider genetic association studies in patient populations

What are the optimal methods for membrane protein extraction when studying C1orf95?

Extraction of membrane proteins like C1orf95 requires specialized approaches:

• Buffer composition optimization:

  • Test multiple detergent classes (non-ionic, zwitterionic, and ionic)

  • Identify optimal detergent concentration for efficient extraction without denaturation

  • Include appropriate protease inhibitors to prevent degradation

  • Optimize salt concentration and pH for maximum stability

• Extraction protocol variables:

  • Temperature control during extraction (typically 4°C to reduce degradation)

  • Incubation time optimization (balance between yield and potential denaturation)

  • Mechanical disruption methods (sonication, homogenization, nitrogen cavitation)

  • Sequential extraction for differential solubilization of membrane compartments

• Sample-specific considerations:

  • For tissue homogenates: additional homogenization may be required compared to cell lysates

  • For cell culture: gentler extraction methods may be sufficient

  • For enriched membrane fractions: specialized detergents like DDM or LMNG may be optimal

• Quality control metrics:

  • Purity assessment through Western blotting for membrane markers

  • Activity/functionality preservation validation when possible

  • Yield quantification using Bradford or BCA assays with detergent-compatible formulations

What methodologies are recommended for studying C1orf95 protein-protein interactions?

For identifying and characterizing C1orf95 interactions:

• Proximity-based methods:

  • BioID fusion protein expression for biotinylation of proximal proteins

  • APEX2 tagging for peroxidase-mediated labeling of neighboring proteins

  • Split-protein complementation assays (BiFC, NanoBiT) for direct interaction testing

• Pull-down approaches:

  • Optimized co-immunoprecipitation protocols with membrane-compatible detergents

  • Tandem affinity purification with careful detergent selection

  • Label-free quantitative proteomics to identify enriched interactors

• Validation strategies:

  • Reverse co-immunoprecipitation with antibodies against identified interactors

  • FRET or BRET analysis for direct protein proximity measurement

  • Functional assays to confirm biological relevance of interactions

• Pathway-focused investigations:

  • Based on expression correlation in muscle tissue contexts, interactions with calcium signaling components may be particularly relevant

  • Components of HIF-1 signaling pathway (such as TEK and NOS3) could be prioritized based on contexts where C1orf95 shows differential expression

How does C1orf95 research fit within the broader context of chromosome 1 studies?

Chromosome 1 harbors approximately 3,000 genes and is associated with numerous disease conditions including Hutchinson-Gilford progeria (LMNA gene), familial adenomatous polyposis (MUTYH gene), Stickler syndrome, Parkinson's disease, Gaucher disease, and Usher syndrome . Additionally, a breakpoint in 1q disrupts the DISC1 gene linked to schizophrenia, and aberrations in chromosome 1 are associated with various cancers including head and neck cancer, malignant melanoma, and multiple myeloma .

As an uncharacterized protein encoded on chromosome 1, C1orf95 research may contribute to our understanding of these disease mechanisms. Systematic characterization of all chromosome 1 proteins, including C1orf95, is essential for comprehensive understanding of genomic function and disease pathophysiology.

What are the most promising directions for future C1orf95 research?

Based on current knowledge gaps and research contexts:

• Functional characterization:

  • Systematic phenotypic analysis of C1orf95 knockout/knockdown models

  • Investigation of potential roles in calcium homeostasis based on expression context

  • Exploration of tissue-specific functions, particularly in muscle and neural tissues

• Structural biology:

  • High-resolution structure determination using cryo-EM or X-ray crystallography

  • Structure-function relationship studies with mutational analysis

  • Ligand binding site identification

• Disease associations:

  • Analysis of C1orf95 expression in patient samples across multiple disease contexts

  • Genetic association studies examining C1orf95 variants in disease cohorts

  • Investigation of potential roles in muscle pathologies based on differential expression data

• Technological development:

  • Creation of improved tools for C1orf95 detection and manipulation

  • Development of conditional knockout models for tissue-specific functional studies

  • Implementation of high-throughput screening approaches to identify C1orf95 modulators