Recombinant Gorilla gorilla gorilla Taste receptor type 2 member 10 (TAS2R10)

What is TAS2R10 and what is its classification within taste receptor families?

TAS2R10 (Taste Receptor Type 2 Member 10) belongs to the TAS2R family of bitter taste receptors. This receptor family is distinct from the TAS1R family, which comprises three genes in most mammals: TAS1R1, TAS1R2, and TAS1R3, responsible for umami and sweet taste perception . The TAS2R gene repertoire is notably larger than the TAS1R gene family, indicating the evolutionary importance of bitter taste detection. TAS2R10 specifically is classified as a broadly tuned bitter receptor, capable of recognizing approximately one-third of bitter components tested thus far .

What is the primary amino acid sequence of Gorilla gorilla gorilla TAS2R10?

The full amino acid sequence of Gorilla gorilla gorilla TAS2R10 is as follows:

mLRVVEGIFIFVVISEXVFGVLGNGFIGLVNCIDCAKNKLSTIGFILTGLAISRIFLIWI IITDGFIQIFSPDIYASGNLIEYISYFWVIGNQSSMWFATSLSIFYFLKIANFSNYIFLW LKSRTNMVLPFMIVFLLISSLLNFAHIAKILNDYKMKNDTVWDLNMYKSEYFIKQILLNL GVIFFFTLSLITCVFLIISLWRHNRQMQSNVTGLRDSNTEAHVKAMKVLISFXILFILYF IGMAIEISCFTVRENKLLLMFGMTTTAIYPWGHSFILILGNSKLKQASLRVLQQLKCCEK RKNLRVT

The expression region spans from amino acid positions 1-307, constituting the full-length protein. The protein's UniProt accession number is Q645Z4, which researchers can use to access additional standardized information about this protein in public databases .

What are the primary tissues where TAS2R10 is expressed?

While TAS2R10 was initially identified in taste buds on the tongue for bitter taste perception, research has revealed that it is widely expressed in multiple extra-oral tissues. The receptor has been confirmed to be mainly expressed in cell lines including HeLa, TPC1, and CAPAN-2, suggesting tissue-specific roles beyond taste sensation . Additionally, TAS2R10 is highly expressed in airway smooth muscle (ASM) cells, with transcript levels 3 to 4-fold higher than those of ADRB2 (which encodes the β2 adrenergic receptor) . Further research has identified significant upregulation of TAS2R10 in lymphocytic populations, indicating potential immunological functions .

What are the optimal storage conditions for recombinant TAS2R10 protein?

For recombinant Gorilla gorilla gorilla TAS2R10 protein, the recommended storage conditions are as follows:

• Short-term storage: Store working aliquots at 4°C for up to one week

• Medium-term storage: Store at -20°C in a storage buffer containing Tris-based buffer with 50% glycerol optimized for this protein

• Long-term storage: For extended preservation, store at -20°C or -80°C

• Important note: Repeated freezing and thawing is not recommended as it may compromise protein integrity and function

These storage recommendations are critical for maintaining the structural integrity and biological activity of the recombinant protein for experimental applications.

How can co-expression analysis be applied to study TAS2R10 functions?

Co-expression analysis represents a powerful bioinformatics approach for elucidating novel functions of TAS2R10. The methodology involves:

• Data collection: Utilizing large-scale transcriptomic datasets, such as the 60,000 Affymetrix expression arrays and 5,000 The Cancer Genome Atlas (TCGA) datasets used in previous research

• Correlation analysis: Identifying genes whose expression patterns positively correlate with TAS2R10 across different tissues and conditions

• Functional enrichment analysis: Applying Gene Ontology (GO) categorization to the co-expressed genes to identify enriched biological processes, cellular components, and molecular functions

• Pathway mapping: Analyzing the enriched genes for common pathways using databases such as KEGG

• Experimental validation: Confirming bioinformatically predicted associations through techniques such as RT-qPCR, as demonstrated in the confirmation of the association between TAS2R10 and ANAPC5 in human thyroid tissue

This comprehensive approach has successfully revealed unexpected roles of TAS2R10 in cellular protein metabolic processes, protein modification processes, and cellular component assembly, providing new research directions beyond simple taste perception .

What are the extra-oral functions of TAS2R10 beyond taste perception?

Research has uncovered several significant biological roles for TAS2R10 beyond its canonical function in bitter taste perception:

• Smooth muscle relaxation: TAS2R10 has been demonstrated to induce relaxation of smooth muscles in multiple tissues:

  • Ileum smooth muscle

  • Airway smooth muscle cells (ASM)

  • Blood vessel smooth muscles

• Cellular processes:

  • Involvement in cellular protein metabolic processes

  • Participation in protein modification processes

  • Cellular component assembly functions

• Molecular activities:

  • Hexosaminidase activity

  • Cytoskeletal adaptor activity

  • Cyclin binding

  • β-N-acetylhexosaminidase activity

• Pathway interactions:

  • Notably, TAS2R10 may be involved in ubiquitin-mediated proteolysis

  • Potential interaction with ANAPC5 (a ubiquitin ligase that controls cell cycle progression)

• Potential tumor-suppressor function:

  • Involvement in cellular processes of cell cycle regulation

  • May exert regulatory functions in cancer biology

• Immune system roles:

  • Significant upregulation in lymphocytic populations

  • Potential involvement in immune response mechanisms

• Disease associations:

  • Suggested linkage to Salmonella infection susceptibility

  • Therapeutic potential in asthma treatment through ASM relaxation

These diverse functions indicate that TAS2R10 has evolved multifunctional capabilities beyond its original role in taste perception, making it an important target for interdisciplinary research.

How does TAS2R10 function in airway smooth muscle and what are its therapeutic implications for asthma?

TAS2R10 exhibits significant expression in airway smooth muscle (ASM) cells, with transcript levels 3-4 fold higher than that of the β2 adrenergic receptor (ADRB2) . This expression pattern has important functional and therapeutic implications:

• Mechanism of action:

  • Activation of TAS2R10 on ASM cells results in robust muscle relaxation

  • This effect is distinct from β2 adrenergic receptor-mediated pathways

  • The signaling cascade likely involves calcium mobilization and cytoskeletal rearrangements

• Experimental evidence:

  • Studies in multiple species including humans, guinea pigs, and mice have confirmed TAS2R expression and function in ASM

  • TAS2R10, along with TAS2R4 and TAS2R14, are particularly important in this context

• Therapeutic potential for asthma:

  • Bronchodilation: TAS2R10 agonists could serve as novel bronchodilators

  • Complementary mechanism: Could work additively with existing β2-agonist therapies

  • Reduced tachyphylaxis: Potential for decreased receptor desensitization compared to β2-agonists

  • Alternative for steroid-resistant patients: Providing new options for difficult-to-treat cases

• Research approaches:

  • Ex vivo tissue studies using precision-cut lung slices

  • In vivo animal models of airway hyperresponsiveness

  • Measurement of airway resistance and compliance

  • Calcium imaging to track intracellular signaling events

This research direction represents a promising avenue for developing new therapeutic approaches for asthma and other obstructive airway diseases, utilizing TAS2R10's natural function in ASM relaxation.

What cellular pathways is TAS2R10 involved in based on co-expression analyses?

Comprehensive bioinformatics analyses of genes positively co-expressed with TAS2R10 have revealed its involvement in several key cellular pathways:

• Protein metabolism and modification pathways:

  • Cellular protein metabolic processes

  • Protein modification processes

  • Cellular protein modification processes

• Cellular structural organization:

  • Cellular component assembly

  • Cytoskeletal organization

  • Association with SAGA-type complexes and SAGA complex

• Enzymatic activity pathways:

  • Hexosaminidase activity pathways

  • β-N-acetylhexosaminidase activity networks

  • Cytoskeletal adaptor activity

• Cell cycle regulation:

  • Cyclin binding pathways

  • Cell cycle progression mechanisms

  • Potential tumor suppression activity

• Ubiquitin pathway:

  • Ubiquitin-mediated proteolysis

  • Interaction with ANAPC5 (a ubiquitin ligase controlling cell cycle progression through ubiquitination)

  • This association was experimentally confirmed in human thyroid tissue via RT-qPCR

• Immune response pathways:

  • Potential involvement in Salmonella infection response

  • Upregulation in lymphocytic populations

These diverse pathway involvements suggest that TAS2R10 serves as a multifunctional receptor that has evolved beyond its original taste-sensing role to participate in fundamental cellular processes across various tissues.

How does TAS2R10 differ between humans and gorillas?

While the search results do not provide comprehensive comparative data between human and gorilla TAS2R10, we can draw several inferences based on available information:

Researchers investigating the evolutionary aspects of TAS2R10 should consider both the conserved functions related to bitter taste perception and the potentially divergent extra-oral functions that might have developed differently between humans and gorillas based on their distinct evolutionary pressures.

How has the TAS2R taste receptor family evolved across primates?

The evolution of taste receptors across primates provides important insights into dietary adaptations and environmental pressures. While the search results focus more on TAS2R10 specifically, we can contextualize its evolution within the broader TAS2R family:

• Evolutionary patterns:

  • Unlike the TAS1R family which was previously thought to be highly conserved, the TAS2R family shows considerable evolutionary plasticity across species

  • This contradicts earlier assumptions that taste genes necessary for nutrient uptake would remain highly conserved

• Phylogenetic relationships:

  • Studies of molecular phylogenetics have used PCR amplification of taste receptor genes (including the design of primers to amplify nucleotides from exons) to establish evolutionary relationships

  • The phylogenetic tree shows that human sweet taste receptor TAS1R2 (a different taste receptor family) is monophyletic with Pan troglodytes and Gorilla gorilla with 100% bootstrap support, suggesting similar patterns might exist for TAS2R receptors

• Adaptive changes:

  • Taste receptor genes undergo major mutations in response to dietary specialization

  • Examples include pseudogenization of TAS1R1 in giant pandas and TAS1R2 in cats and vampire bats

  • Similar adaptive changes likely occur in the TAS2R family based on dietary preferences

• Research approaches:

  • PCR amplification and sequencing of taste receptor genes from diverse primate species

  • Phylogenetic analysis to determine evolutionary relationships

  • Functional assays to correlate genetic changes with altered receptor function

  • Correlation of genetic changes with known dietary adaptations

Understanding the evolution of TAS2R10 within the context of primate evolution provides valuable insights into how taste perception has shaped dietary preferences and how, conversely, dietary specialization has driven taste receptor evolution.

What are the optimal methods for studying TAS2R10 function in different tissue types?

Investigating TAS2R10 function across diverse tissues requires tissue-specific experimental approaches:

• Airway smooth muscle (ASM) studies:

  • Ex vivo precision-cut lung slices to measure contractile responses

  • Calcium imaging to detect intracellular signaling

  • Measurement of ASM cell relaxation in response to TAS2R10 agonists

  • Comparison with standard bronchodilators like β2-agonists

• Immune cell investigations:

  • Flow cytometry to identify TAS2R10-expressing lymphocyte populations

  • Functional assays to determine receptor activation effects on immune cell function

  • RT-qPCR to quantify expression levels in different immune cell subsets

• Cell line-based studies:

  • Use of HeLa, TPC1, and CAPAN-2 cell lines which have confirmed TAS2R10 expression

  • siRNA knockdown or CRISPR-Cas9 genome editing to analyze loss-of-function effects

  • Overexpression studies to assess gain-of-function effects

• Protein-protein interaction studies:

  • Co-immunoprecipitation to confirm interactions with proteins like ANAPC5

  • Bioluminescence resonance energy transfer (BRET) or fluorescence resonance energy transfer (FRET) to study protein interactions in living cells

  • Proximity ligation assays to detect protein interactions in tissue samples

• Signal transduction analysis:

  • Measurement of second messengers (calcium, cAMP) following receptor activation

  • Western blotting to assess phosphorylation of downstream effectors

  • Reporter gene assays to quantify transcriptional responses

These methodological approaches provide complementary data to understand the tissue-specific functions of TAS2R10 beyond its canonical role in taste perception.

How can researchers effectively validate bioinformatically predicted functions of TAS2R10?

Validating bioinformatically predicted functions requires a systematic approach combining multiple experimental strategies:

• Expression correlation validation:

  • RT-qPCR to confirm co-expression of TAS2R10 with predicted interaction partners

  • This approach was successfully used to validate the association between TAS2R10 and ANAPC5 in human thyroid tissue

• Protein-protein interaction confirmation:

  • Co-immunoprecipitation (Co-IP) experiments

  • Proximity ligation assays

  • Yeast two-hybrid assays

  • Bioluminescence/fluorescence resonance energy transfer (BRET/FRET)

• Functional pathway validation:

  • For ubiquitin-mediated proteolysis involvement:

    • Measure ubiquitination states of target proteins with and without TAS2R10 manipulation

    • Assess proteasome activity in TAS2R10-expressing versus non-expressing cells

    • Determine effects of proteasome inhibitors on TAS2R10-mediated functions

• Cell-based functional assays:

  • For smooth muscle relaxation:

    • Measure muscle tension in tissue preparations

    • Calcium imaging to track signaling events

    • Assessment of cytoskeletal rearrangements

  • For cell cycle regulation:

    • Flow cytometry to analyze cell cycle distribution

    • BrdU incorporation to measure proliferation

    • Analysis of cyclin levels and activity

• Disease model validation:

  • For Salmonella infection association:

    • Infection assays in cell culture with TAS2R10 manipulation

    • Assessment of bacterial invasion and replication

    • Analysis of host immune responses

  • For asthma therapeutic potential:

    • Measurement of bronchodilation in ex vivo lung slices

    • In vivo assessment in animal models of airway hyperresponsiveness

By systematically applying these validation approaches, researchers can confirm bioinformatically predicted functions and establish the biological relevance of TAS2R10 in various physiological and pathological contexts.