苦味受体与甜味、鲜味受体具有不同的进化途径

通过生物信息学和系统发育学分析,研究了苦味受体和甜味／鲜味受体的进化途径。结果显示,苦味受体和甜味／鲜味受体在进化上具有远相关,并且具有不同的进化途径,提示这可能是导致这些受体具有不同功能,传导不同味觉的原因。     

The human bitter taste receptor hTAS2R39 is the primary receptor for the bitterness of theaflavins

We purified several hundred mgs of four major theaflavins (theaflavin, theaflavin-3-O-gallate, theaflavin-3′-O-gallate, and theaflavin-3,3′-O-digallate). Among the 25 hTAS2Rs expressed in HEK293T cells, hTAS2R39 and hTAS2R14 were activated by theaflavins. Both hTAS2R39 and hTAS2R14 responded to theaflavin-3′-O-gallate. In addition, hTAS2R39 was activated by theaflavin and theaflavin-3,3′-O-gallate, but not by theaflavin-3-O-gallate. In contrast, hTAS2R14 responded to theaflavin-3-O-gallate.     

6-Methoxyflavanones as Bitter Taste Receptor Blockers for hTAS2R39

Many (dietary) bitter compounds, e.g. flavonoids, activate bitter receptor hTAS2R39 in cell-based assays. Several flavonoids, amongst which some flavanones, are known not to activate this receptor. As certain flavanones are known to mask bitter taste sensorially, flavanones might act as bitter receptor antagonists. Fourteen flavanones were investigated for their potential to reduce activation of hTAS2R39 by epicatechin gallate (ECG), one of the main bitter compounds occurring in green tea. Three flavanones showed inhibitory behavior towards the activation of hTAS2R39 by ECG: 4′-fluoro-6-methoxyflavanone, 6,3′-dimethoxyflavanone, and 6-methoxyflavanone (in order of decreasing potency). The 6-methoxyflavanones also inhibited activation of hTAS2R14 (another bitter receptor activated by ECG), though to a lesser extent. Dose-response curves of ECG at various concentrations of the full antagonist 4′-fluoro-6-methoxyflavanone and wash-out experiments indicated reversible insurmountable antagonism. The same effect was observed for the structurally different agonist denatonium benzoate.     

Human bitter taste receptors hTAS2R8 and hTAS2R39 with differential functions to recognize bitter peptides

The strong bitter peptide, Phe-Phe-Pro-Arg, activated cultured cells expressing either of the known human bitter taste receptors, hTAS2R8 and hTAS2R39. The partial structure of Pro-Arg activated hTAS2R39, but did not activate hTAS2R8. These receptors may not indiscriminately recognize bitter peptides, but have a differential function in their recognition.     

黄芩苷通过苦味受体促进哮喘小鼠呼吸道炎性细胞凋亡

全球哮喘患者有3亿多人。目前,约有一半患者的病情不能较好地用现有药物来控制。因此,寻找新的更有效的治疗哮喘病的药物是非常必要的。最近的研究发现,苦味受体(bitter taste receptors,Tas2rs)在呼吸系统中表达,且苦味剂对哮喘有治疗潜力,苦味受体可能成为哮喘治疗的新靶点。为此,本文研究了苦味化合物黄芩苷(baicalin,BA)对哮喘的干预作用,分析黄芩苷对哮喘小鼠呼吸道炎性细胞凋亡的干预作用及其与苦味信号转导的关系。选雄性BALB/c小鼠,随机分为对照组(CK组)、腹腔注射致敏加雾化吸入卵清蛋白(ovalbumin,OVA)激发制成的哮喘模型组(OVA组)和黄芩苷灌胃干预哮喘组(OVA+BA组)。结果发现,OVA组小鼠肺泡灌洗液中白细胞总数和分类细胞计数显著增加,黄芩苷干预组白细胞数量显著减少;HE染色后,OVA组小鼠肺组织中可见炎性细胞浸润、肺泡隔增厚和肺泡囊缩小,上述症状在OVA+BA组小鼠肺部明显减轻;实时荧光定量RT-PCR检测发现,肺组织中黏蛋白Muc5ac表达水平在OVA组明显增高(P <0.05),黄芩苷干预组显著低于OVA组(P <0.05)。OVA致敏哮喘小鼠呼吸道中Tas2r108、Tas2r126、Tas2r135和Tas2r143及其下游信号转导分子α-gust和Trpm5下调表达(P <0.05),促凋亡因子P53、Bax和胱天蛋白酶(caspase,Casp)Casp3转录抑制,凋亡抑制基因Bcl2上调表达,胱天蛋白酶3活性显著降低(P <0.05);黄芩苷干预组4个Tas2rs及苦味信号转导分子转录上调(P <0.05),促凋亡基因P53、Bax和Casp3转录上调,Bcl2转录抑制,胱天蛋白酶3活性显著高于OVA组(P <0.05)。结果表明,黄芩苷干预可激活哮喘鼠呼吸道苦味信号转导通路,并使呼吸道炎性细胞减少、黏蛋白分泌减少。即苦味物质黄芩苷可能作为一种苦味受体激动剂,通过激活苦味信号转导系统促进呼吸道炎性细胞凋亡,减轻肺部炎症和损伤,缓解哮喘发作。     

Bitter peptides activate hTAS2Rs, the human bitter receptors

Fermented food contains numerous peptides derived from material proteins. Bitter peptides formed during the fermentation process are responsible for the bitter taste of fermented food. We investigated whether human bitter receptors (hTAS2Rs) recognize bitterness of peptides with a heterologous expression system. HEK293 cells expressing hTAS2R1, hTAS2R4, hTAS2R14, and hTAS2R16 responded to bitter casein digests. Among those cells, the hTAS2R1-expressing cell was most strongly activated by the synthesized bitter peptides Gly-Phe and Gly-Leu, and none of the cells was activated by the non-bitter dipeptide Gly-Gly. The results showed that these bitter peptides, as well as many other bitter compounds, activate hTAS2Rs, suggesting that humans utilize these hTAS2Rs to recognize and perceive the structure and bitterness of peptides.     

Local Adaptation of Bitter Taste and Ecological Speciation in a Wild Mammal

Sensory systems are attractive evolutionary models to address how organisms adapt to local environments that can cause ecological speciation. However, tests of these evolutionary models have focused on visual, auditory, and olfactory senses. Here, we show local adaptation of bitter taste receptor genes in two neighboring populations of a wild mammal—the blind mole rat Spalax galili—that show ecological speciation in divergent soil environments. We found that basalt-type bitter receptors showed higher response intensity and sensitivity compared with chalk-type ones using both genetic and cell-based functional analyses. Such functional changes could help animals adapted to basalt soil select plants with less bitterness from diverse local foods, whereas a weaker reception to bitter taste may allow consumption of a greater range of plants for animals inhabiting chalk soil with a scarcity of food supply. Our study shows divergent selection on food resources through local adaptation of bitter receptors, and suggests that taste plays an important yet underappreciated role in speciation.     

呼吸道苦味信号转导抑制与小鼠哮喘相关

鉴于哮喘病患病人数众多,约有一半的病人病情得不到较好的控制,急需新的治疗方法和药物.最近研究发现,苦味受体(bitter taste receptors,T2 Rs)在多个组织中表达,且苦味剂对哮喘有治疗潜力,T2Rs有可能成为哮喘治疗的新靶点.本文选C57BL/6小鼠随机分为对照组、二氧化硫(sulfur dioxi...     

Snooker Structure-Based Pharmacophore Model Explains Differences in Agonist and Blocker Binding to Bitter Receptor hTAS2R39

The human bitter taste receptor hTAS2R39 can be activated by many dietary (iso)flavonoids. Furthermore, hTAS2R39 activity can be blocked by 6-methoxyflavanones, 4’-fluoro-6-methoxyflavanone in particular. A structure-based pharmacophore model of the hTAS2R39 binding pocket was built using Snooker software, which has been used successfully before for drug design of GPCRs of the rhodopsin subfamily. For the validation of the model, two sets of compounds, both of which contained actives and inactives, were used: (i) an (iso)flavonoid-dedicated set, and (ii) a more generic, structurally diverse set. Agonists were characterized by their linear binding geometry and the fact that they bound deeply in the hTAS2R39 pocket, mapping the hydrogen donor feature based on T5.45 and N3.36, analogues of which have been proposed to play a key role in activation of GPCRs. Blockers lack hydrogen-bond donors enabling contact to the receptor. Furthermore, they had a crooked geometry, which could sterically hinder movement of the TM domains upon receptor activation. Our results reveal characteristics of hTAS2R39 agonist and bitter blocker binding, which might facilitate the development of blockers suitable to counter the bitterness of dietary hTAS2R39 agonists in food applications.     

Top-Down Control of Sweet and Bitter Taste in the Mammalian Brain

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Functional diversity of bitter taste receptor TAS2R16 in primates

In mammals, bitter taste is mediated by TAS2R genes, which belong to the large family of seven transmembrane G protein-coupled receptors. Because TAS2Rs are directly involved in the interaction between mammals and their dietary sources, it is likely that these genes evolved to reflect species-specific diets during mammalian evolution. Here, we investigated the sensitivities of TAS2R16s of various primates by using a cultured cell expression system, and found that the sensitivity of each primate species varied according to the ligand. Especially, the sensitivity of TAS2R16 of Japanese macaques to salicin was much lower than that of human TAS2R16, which was supported by behavioural tests. These results suggest the possibility that bitter-taste sensitivities evolved independently by replacing specific amino acid residues of TAS2Rs in different primate species to adapt to food items they use.     

羚牛苦味受体3(Tas2R3)基因的克隆及生物信息学分析

Tas2R3是苦味受体基因家族中一个重要的成员,为了进一步了解和研究羚牛(Budorcas taxicolor)苦味受体基因的结构和功能,本研究对羚牛苦味受体3 (Tas2R3)基因进行了克隆和生物信息学分析(GenBank登录号:MG650195)。结果显示,羚牛Tas2R3基因编码区(coding sequence, CDS)序列全长951 bp,共编码316个氨基酸,以亮氨酸含量最高,谷氨酰胺含量最低。其蛋白质等电点为9.68,分子量为51.96 kD。高级结构功能预测显示,二级结构以α-螺旋为主,蛋白质为碱性、稳定的亲水性蛋白,由4个胞外区、7个跨膜区和4个胞内区组成。预测到2种类型共8个糖基化功能位点和4种类型共15个磷酸化功能位点。通过比较Tas2R3基因种间相似性发现,在偶蹄目中具有很高的同源性,羚牛与绵羊(Ovis aries)的相似性最高(0.98),与褐家鼠(Rattus norvegicus)最低(0.52)。用羚牛、绵羊等12个物种的Tas2R3基因CDS序列构建的NJ树与ME树结构一致,表明Tas2R3基因适合用于构建不同物种间的系统进化树。     

Association of Bitter Taste Receptor T2R38 Polymorphisms,Oral Microbiota,and Rheumatoid Arthritis

The association of taste genetics and the oral microbiome in autoimmune diseases such as rheumatoid arthritis (RA) has not been reported. We explored a novel oral mucosal innate immune pathway involving the bitter taste G protein-coupled receptor T2R38. This case–control study aimed to evaluate whether T2R38 polymorphisms associate with the buccal microbial composition in RA. Genomic DNA was obtained from buccal swabs of 35 RA patients and 64 non-RA controls. TAS2R38 genotypes were determined by Sanger sequencing. The buccal microbiome was assessed by Illumina MiSeq sequencing of the V4-16S rRNA gene. Bacterial community differences were analyzed with alpha and beta diversity measures. Linear discriminant analysis effect size identified taxa discriminating between RA versus non-RA and across TAS2R38 genotypes. TAS2R38 genotype frequency was similar between RA and non-RA controls (PAV/PAV; PAV/AVI; AVI/AVI: RA 42.9%; 45.7%; 11.4% versus controls 32.8%; 48.4%; 18.8%, chi-square (2, N = 99) = 2.1, p = 0.35). The relative abundance of Porphyromonas, among others, differed between RA and non-RA controls. The relative abundance of several bacterial species also differed across TAS2R38 genotypes. These findings suggest an association between T2R38 polymorphisms and RA buccal microbial composition. However, further research is needed to understand the impact of T2R38 in oral health and RA development.     

Evaluation of the bitterness of green tea catechins by a cell-based assay with the human bitter taste receptor hTAS2R39

Catechins have a broad range of physiological functions and act as the main taste ingredient of green tea. Although catechins show a strong bitterness, the bitter taste receptor for catechins has not been fully understood. The objective of this study was to identify the receptor for the major green tea catechins such as (−)-epicatechin (EC), (−)-epicatechin gallate (ECg), (−)-epigallocatechin (EGC), and (−)-epigallocatechin gallate (EGCg). By the cell-based assay using cultured cells expressing human bitter taste receptor, a clear response of hTAS2R39-expressing cells was observed to 300 μM of either ECg or EGCg, which elicit a strong bitterness in humans. The response of hTAS2R39-expressing cells to ECg was the strongest among the tested catechins, followed by EGCg. Because the cellular response to EC and EGC is much weaker than those of ECg and EGCg, galloyl groups was strongly supposed to be involved in the bitter intensity. This finding is similar to the observations of taste intensity obtained from a human sensory study. Our results suggest the participation of hTAS2R39 in the detection of catechins in humans, indicating the possibility that bitterness of tea catechins can be evaluated by using cells expressing hTAS2R39.     

A New World Monkey Resembles Human in Bitter Taste Receptor Evolution and Function via a Single Parallel Amino Acid Substitution

Bitter taste receptors serve as a vital component in the defense system against toxin intake by animals, and the family of genes encoding these receptors has been demonstrated, usually by family size variance, to correlate with dietary preference. However, few systematic studies of specific Tas2R to unveil their functional evolution have been conducted. Here, we surveyed Tas2R16 across all major clades of primates and reported a rare case of a convergent change to increase sensitivity to β-glucopyranosides in human and a New World monkey, the white-faced saki. Combining analyses at multiple levels, we demonstrate that a parallel amino acid substitution (K172N) shared by these two species is responsible for this functional convergence of Tas2R16. Considering the specialized feeding preference of the white-faced saki, the K172N change likely played an important adaptive role in its early evolution to avoid potentially toxic cyanogenic glycosides, as suggested for the human TAS2R16 gene.     

Bitter taste perception in Neanderthals through the analysis of the TAS2R38 gene

The bitter taste perception (associated with the ability or inability to taste phenylthiocarbamide) is mediated by the TAS2R38 gene. Most of the variation in this gene is explained by three common amino-acid polymorphisms at positions 49 (encoding proline or alanine), 262 (alanine or valine) and 296 (valine or isoleucine) that determine two common isoforms: proline–alanine–valine (PAV) and alanine–valine–isoleucine (AVI). PAV is the major taster haplotype (heterozygote and homozygote) and AVI is the major non-taster haplotype (homozygote). Amino acid 49 has the major effect on the distinction between tasters and non-tasters of all three variants. The sense of bitter taste protects us from ingesting toxic substances, present in some vegetables, that can affect the thyroid when ingested in large quantities. Balancing selection has been used to explain the current high non-taster frequency, by maintaining divergent TAS2R38 alleles in humans. We have amplified and sequenced the TAS2R38 amino acid 49 in the virtually uncontaminated Neanderthal sample of El Sidrón 1253 and have determined that it was heterozygous. Thus, this Neanderthal was a taster individual, although probably slightly less than a PAV homozygote. This indicates that variation in bitter taste perception pre-dates the divergence of the lineages leading to Neanderthals and modern humans.     

TAS2R20 variants confer dietary adaptation to high‐quercitrin bamboo leaves in Qinling giant pandas

Sensitivity to bitter tastes provides animals with an important means of interacting with their environment and thus, influences their dietary preferences. Genetic variants encoding functionally distinct receptor types contribute to variation in bitter taste sensitivity. Our previous study showed that two nonsynonymous sites, A52V and Q296H, in the TAS2R20 gene are directionally selected in giant pandas from the Qinling Mountains, which are speculated to be the causative base‐pair changes of Qinling pandas for the higher preference for bamboo leaves in comparison with other pandas. Here, we used functional expression in engineered cells to identify agonists of pTAS2R20 (i.e., giant panda's TAS2R20) and interrogated the differences in perception in the in vitro responses of pTAS2R20 variants to the agonists. Our results show that pTAS2R20 is specifically activated by quercitrin and that pTAS2R20 variants exhibit differences in the sensitivity of their response to the agonist. Compared with pTAS2R20 in pandas from other areas, the receptor variant with A52V and Q296H, which is most commonly found in Qinling pandas, confers a significantly decreased sensitivity to quercitrin. We subsequently quantified the quercitrin content of the leaves of bamboo distributed in the Qinling Mountains, which was found to be significantly higher than that of the leaves of bamboo from panda habitats in other areas. Our results suggest that the decreased sensitivity to quercitrin in Qinling pandas results in higher‐quercitrin‐containing bamboo leaves to be tasting less bitter to them and thus, influences their dietary preference. This study illustrates the genetic adaptation of Qinling pandas to their environments and provides a fine example of the functional effects of directional selection in the giant panda.     

Bitter taste thresholds, numbers and diameters of circumvallate papillae and their relation with age in a Turkish population

Objectives: This study analyses the relationship between the sense of bitter taste and age. The relationships between these and the numbers and diameters of Circumvallate Papillae (CP) are also analysed. Subjects: Twenty-four elderly subjects (from 65 to 85 years) and 30 young subjects (from 17 to 25 years) were studied. Method: Bitter taste thresholds were determined by the three drop method with an ascending series of concentrations. The numbers and diameters of CP were observed by direct naked eye observation using a wooden tongue depressor, a gauge and a light source. Results: The bitter taste acuity was significantly poorer in the older sample. There was no relationship between the bitter taste acuity and number of papillae but an inverse relationship was observed between the bitter taste acuity and diameters of papillae, thus lower acuity was associated with larger papillae. It was also observed that the diameters of papillae were inversely related to the numbers of CP. The distribution of numbers and diameters of CP were not significantly different between these samples with age.     

猪獾苦味受体T2R2基因的分子克隆与进化分析

苦味的感知是机体有效的自我保护机制之一。文章采用PCR和克隆测序方法首次从猪獾基因组中获得一全长为1 169 bp的苦味受体T2R2基因DNA序列(GenBank登录号: FJ812727)。该序列含有完整的1个外显子(无内含子), 大小为915 bp, 编码304个氨基酸残基。其蛋白质等电点为9.76, 分子量为34.74 kDa。拓扑结构预测显示猪獾T2R2蛋白上含有N-糖基化位点、N-肉豆蔻酰化位点各1个, 蛋白激酶C磷酸化位点2个。整个蛋白质多肽链含有7个跨膜螺旋区, 4个细胞外区和4个细胞内区。亲水性/疏水性分析表明, 猪獾T2R2蛋白质为一疏水性蛋白, 其亲水性区段所占比例较小。种间相似性比较显示, 猪獾T2R2基因与犬、猫、牛、马、黑猩猩和小鼠的T2R2基因cDNA序列相似性分别为91.4%、90.6%、84.4%、85.4%、83.8%、72.1%, 氨基酸序列相似性分别为85.5%、85.8%、74.0%、77.6%、75.3%、61.5%。核苷酸替换计算和选择性检验结果表明, 猪獾T2R2基因与犬、猫、牛、马、黑猩猩和小鼠间存在着强烈的纯净化选择(Purifying selection), 即强烈的功能束缚(Functional constraint), 进一步分析发现该选择作用实际上主要存在于跨膜区。猪獾、犬、猫、牛、马、黑猩猩和小鼠的T2R2基因外显子核苷酸序列构建的基因树与其物种树的拓扑结构是相一致的, 表明T2R2基因适合于构建不同物种间的系统进化树。