Recombinant Human Transmembrane protein 183B (TMEM183B)

What is TMEM183B and what is its cellular localization?

TMEM183B (also known as C1ORF37DUP) is a transmembrane protein associated with cell membranes that may be involved in cell-cell or cell-environment interactions. The protein contains a complete open reading frame that has been demonstrated to be transcribed in a limited number of human tissues . While initially thought to be a pseudogene of chromosome 1 open reading frame 37 due to being intronless and retaining a polyA tail at the 3' end, subsequent research has confirmed its expression and functional significance .

What structural features characterize TMEM183B protein?

TMEM183B is predicted to contain transmembrane domains that anchor it within cellular membranes. For structural analysis, techniques such as those employed for related transmembrane proteins can be applied. Alphafold combined with MMSeq2 can be used to generate 3D structures, with quality assessment via PLDDT and PAE scores . When analyzing transmembrane proteins, it's common practice to truncate loop regions that don't meet desired criteria for cross-validation of structural predictions .

How does TMEM183B differ from other TMEM family proteins?

While TMEM183B shares structural similarities with other transmembrane proteins, it has distinct roles in cellular processes. It's important to note that TMEM183B should not be confused with TMEM184B, which has been associated with neurodevelopmental disorders , or TMEM18, which has been linked to obesity risk . These proteins, despite similar nomenclature, have different functions and disease associations.

What is the role of TMEM183B in cancer progression?

Research has identified TMEM183B as having oncogenic properties in hypopharyngeal squamous cell carcinoma (HPSCC). Studies demonstrate that TMEM183B promotes HPSCC cell growth, invasion, and migration in FaDu cells, while inhibiting cell apoptosis . The expression of TMEM183B is significantly higher in HPSCC tissues compared to adjacent normal tissues, suggesting its potential as an oncogenic driver .

What methodologies are most effective for studying TMEM183B in cancer models?

For investigating TMEM183B in cancer, researchers typically employ:

• Immunohistochemistry (IHC) to determine expression differences between tumor tissues and adjacent normal tissues

• Bioinformatics analysis using The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases to verify expression patterns

• In vitro assays for cell proliferation, invasion, and migration

• Flow cytometry to assess effects on apoptosis

• In vivo experiments to evaluate tumor growth

These methodologies provide comprehensive insights into TMEM183B's oncogenic functions in cancer models.

Can TMEM183B serve as a diagnostic biomarker for specific cancers?

Evidence suggests that TMEM183B might serve as a potential diagnostic biomarker for HPSCC. Research has revealed significantly higher expression of TMEM183B in HPSCC tissues compared to adjacent normal tissues, which has been confirmed through bioinformatics analysis of TCGA and GEO databases . The protein's role in promoting HPSCC cell growth, invasion, and migration while inhibiting apoptosis makes it a promising candidate for both diagnosis and as a therapeutic target .

What expression systems are optimal for recombinant TMEM183B production?

For recombinant TMEM183B production, several expression systems have been utilized for transmembrane proteins. Recombinant TMEM183B protein is available for multiple species including Human, Cynomolgus/Rhesus macaque, Rat, Mouse, Feline, Canine, Bovine, and Equine . While specific expression systems for TMEM183B are not detailed in the search results, researchers commonly use:

• Mammalian cell lines (like HEK293T) for proper post-translational modifications

• Gateway-compatible open reading frame (ORF) expression systems

• In vitro transcription using mMessage mMachine kits for mRNA production

The choice of expression system should align with the intended experimental application.

What are the challenges in working with recombinant transmembrane proteins like TMEM183B?

Transmembrane proteins present unique challenges in research settings:

• Maintaining proper protein folding and stability

• Ensuring correct insertion into membranes during expression

• Preserving functionality when removed from native membrane environments

• Managing protein aggregation during purification

• Achieving sufficient yield for experimental applications

These challenges require specialized approaches in buffer selection, purification methods, and storage conditions to maintain protein integrity.

How can researchers verify the functionality of recombinant TMEM183B?

Functional verification of recombinant TMEM183B can be performed through:

• Cell-based assays measuring proliferation, migration, and invasion capabilities

• Flow cytometry to assess effects on apoptosis in appropriate cell lines

• Structural integrity confirmation using techniques like circular dichroism

• In vivo complementation studies to test protein function

• Cell localization studies using fluorescently tagged protein variants

These approaches provide comprehensive validation of recombinant TMEM183B functionality.

What are appropriate cell lines for studying TMEM183B function?

Based on available research, appropriate cell lines include:

• FaDu cells - hypopharyngeal squamous carcinoma cell line used in TMEM183B cancer research

• HEK293T cells - commonly used for protein expression and cellular localization studies

When selecting cell lines, researchers should consider:

• Endogenous expression levels of TMEM183B

• Tissue relevance to the research question

• Transfection efficiency

• Growth characteristics and handling requirements

How can TMEM183B variants be generated for functional studies?

TMEM183B variants can be generated through:

• Site-directed mutagenesis on Gateway-compatible ORF entry vectors

• Transcript verification through sequencing

• In vitro transcription using appropriate kits (e.g., mMessage mMachine SP6 Transcription kit)

• Linearization of constructs with appropriate restriction enzymes (e.g., NotI)

For variant testing, researchers commonly inject defined amounts (e.g., 200 pg) of mRNA into model systems or transfect cell lines with plasmid constructs encoding the variants .

What in vivo models are suitable for TMEM183B research?

While specific models for TMEM183B are not detailed in the search results, zebrafish has been employed for studying related transmembrane proteins. This model allows for:

• Morpholino-mediated gene suppression

• mRNA injection for overexpression or rescue experiments

• Assessment of developmental phenotypes

• Evaluation of neural development and commissural neurons

• Testing of both dominant and loss-of-function variant hypotheses

Selection of appropriate in vivo models should align with specific research questions and available resources.

How can structural modeling inform TMEM183B variant analysis?

Advanced structural modeling can:

• Predict how variants affect protein stability

• Identify critical functional domains (such as pore domains in related transmembrane proteins)

• Map disease-associated variants to specific structural features

• Guide site-directed mutagenesis experiments

• Assist in developing targeted therapeutic approaches

For structural prediction, tools like Alphafold with MMSeq2 can generate models that are then quality-assessed via PLDDT and PAE scores. The Ramachandran plot provides validation of predicted protein geometry .

What cellular pathways might be regulated by TMEM183B?

While specific pathways for TMEM183B are not fully characterized in the available research, studies on related proteins suggest potential involvement in:

• Cell proliferation, migration, and invasion pathways relevant to cancer progression

• Apoptotic signaling pathways

• Cell-cell or cell-environment interaction pathways

Additionally, related transmembrane proteins have been implicated in cellular metabolic regulation and nutrient signaling pathways including TFEB (transcription factor EB) localization, a master regulator of lysosomal biogenesis .

How can high-throughput approaches advance TMEM183B research?

Modern high-throughput approaches that could advance TMEM183B research include:

• CRISPR-Cas9 screening to identify synthetic lethal interactions or regulatory elements

• Transcriptomics to determine global gene expression changes following TMEM183B modulation

• Proteomics to identify protein interaction networks

• High-content imaging to simultaneously assess multiple cellular phenotypes

• Patient-derived organoids to study TMEM183B in clinically relevant systems

These approaches can provide comprehensive insights into TMEM183B function beyond traditional single-gene studies.

TMEM183B Protein Variants and Their Effects

Available Recombinant TMEM183B Protein Resources

Data compiled from product information