公开(公告)号：US07344859B2

公开(公告)日：2008-03-18

申请号：US10725472

申请日：2003-12-03

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： C12P21/06 ,  C12N1/20 ,  C12N15/74

CPC分类号： G01N33/5041 ,  A01K2217/05 ,  C07K14/705 ,  C07K2319/00 ,  C12Q1/6876 ,  C12Q2600/124 ,  C12Q2600/158 ,  G01N33/5008 ,  G01N33/566 ,  G01N2333/4719 ,  G01N2333/705 ,  G01N2333/726 ,  G01N2500/04

摘要： Methods for expressing a functional heteromeric taste receptor that responds to sweet taste stimuli are provided. These methods comprise the co-expression of T1R2 and T1R3 nucleic acid sequences in a host cell that desirably further expresses a G protein that couples therewith, e.g., Gα15, Gα16 or gustducin. In preferred embodiments, the host cells will be mammalian cells or Xenopus oocytes. These nucleic acid sequences are expressed constitutively or under inducible conditions. In preferred embodiments the expression methods will use HEK-293 cells that also stably express Gα15. These methods give rise to heteromeric receptors and compositions containing that are useful in assays for identifying novel sweeteners and sweetness modulators.

摘要翻译： 提供了表达响应甜味刺激的功能性异味味觉受体的方法。 这些方法包括在宿主细胞中共同表达T1R2和T1R3核酸序列，其希望进一步表达与其偶合的G蛋白，例如Gαααα，αα16 或外排。 在优选的实施方案中，宿主细胞将是哺乳动物细胞或非洲爪蟾卵母细胞。 这些核酸序列组成型或在可诱导条件下表达。 在优选的实施方案中，表达方法将使用也稳定表达Gα1515的HEK-293细胞。 这些方法引起异聚受体，并且含有这些方法的组合物可用于鉴定新型甜味剂和甜味调节剂的测定中。

公开(公告)号：US07309577B2

公开(公告)日：2007-12-18

申请号：US10725418

申请日：2003-12-03

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： G01N33/53 ,  G01N33/567

CPC分类号： C07K14/705 ,  A01K2217/05 ,  G01N2333/726

摘要： Binding assays for identifying compounds that induce or modulate the T1R1/T1R3 (umami) receptor-associated taste are provided. These binding assays detect the specific binding of a compound to a T1R1/T1R3 (umami) taste receptor or detect the modulation (inhibition or enhancement) of the binding of another compound, e.g., L-glutamate, L-aspartate or lactisole to a T1R1/T1R3 umami taste receptor. Compounds that are identified in these binding assays have potential application as T1R1/T1R3 umami taste modulators and therefore can be used as flavor additives in compositions for human or animal consumption.

摘要翻译： 提供了用于鉴定诱导或调节T1R1 / T1R3（鲜味）受体相关味道的化合物的结合测定法。 这些结合测定法检测化合物与T1R1 / T1R3（鲜味）味觉受体的特异性结合，或检测另一种化合物（例如L-谷氨酸，L-天冬氨酸或乳清蛋白）与T1R1的结合的调节（抑制或增强） / T1R3鲜味受体。 在这些结合测定中鉴定的化合物具有作为T1R1 / T1R3鲜味调节剂的潜在应用，因此可用作人或动物消费组合物中的风味添加剂。

公开(公告)号：US07294474B2

公开(公告)日：2007-11-13

申请号：US10725475

申请日：2003-12-03

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： G01N33/53 ,  G01N33/567 ,  C07K1/00 ,  C07K14/00 ,  C07K17/00

CPC分类号： G01N33/5041 ,  A01K2217/05 ,  C07K14/705 ,  C07K2319/00 ,  C12Q1/6876 ,  C12Q2600/124 ,  C12Q2600/158 ,  G01N33/5008 ,  G01N33/566 ,  G01N2333/4719 ,  G01N2333/705 ,  G01N2333/726 ,  G01N2500/04

摘要： Functional assays for identifying compounds that activate or modulate the activation of the T1R2/T1R3 (sweet) taste receptor are provided. These assays detect the effect of one or more compounds on the activation of T1R2/T1R3 (sweet) taste receptor or on the activation of T1R2/T1R3 sweet taste receptor by another, compound e.g., saccharin or another artificial or natural sweetener. These assays preferably are cell-based functional assays and typically use cells, e.g., HEK-293 cells that stably express a G protein such as Gα15, Gα16 or gustducin. Compounds identified in the disclosed functional assays are potentially useful as additives in compositions for human or animal consumption.

摘要翻译： 提供用于鉴定激活或调节T1R2 / T1R3（甜味）味觉受体的活化的化合物的功能测定。 这些测定法检测一种或多种化合物对T1R2 / T1R3（甜味）感受器的激活或另一种化合物例如糖精或另一种人造或天然甜味剂对T1R2 / T1R3甜味受体的活化的影响。 这些测定优选是基于细胞的功能测定，并且通常使用细胞，例如稳定表达G蛋白如GαA，αα16或gustducin的HEK-293细胞。 在所公开的功能测定中鉴定的化合物潜在地用作用于人或动物消费的组合物中的添加剂。

公开(公告)号：US20130102010A1

公开(公告)日：2013-04-25

申请号：US13615778

申请日：2012-09-14

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： G01N33/50

CPC分类号： G01N33/5008 ,  C12N2503/00 ,  C12N2510/00 ,  C40B30/04 ,  G01N33/6872 ,  G01N2333/726 ,  G01N2500/10

摘要： The present invention relates to the discovery that the T1R receptors assemble to form functional taste receptors. Particularly, it has been discovered that co-expression of T1R1 and T1R3 results in a taste receptor that responds to umami taste stimuli, including monosodium glutamate. Also, it has been discovered that co-expression of the T1R2 and T1R3 receptors results in a taste receptor that responds to sweet taste stimuli including naturally occurring and artificial sweeteners.Also the present invention relates to the use of hetero-oligomeric taste receptors comprising T1R1/T1R3 and T1R2/T1R3 in assays to identify compounds that respectively respond to umami taste stimuli and sweet taste stimuli.

摘要翻译： 本发明涉及T1R受体组装形成功能性味觉受体的发现。 特别地，已经发现，T1R1和T1R3的共表达导致对鲜味刺激（包括谷氨酸钠）起反应的味觉感受器。 此外，已经发现，T1R2和T1R3受体的共表达导致对包括天然存在的和人造甜味剂的甜味刺激有反应的味觉受体。 本发明还涉及在测定中使用包含T1R1 / T1R3和T1R2 / T1R3的异低聚味觉感受器来鉴定分别响应于鲜味刺激和甜味刺激的化合物。

公开(公告)号：US08097426B2

公开(公告)日：2012-01-17

申请号：US12270855

申请日：2008-11-14

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： G01N33/567 ,  C07K1/00 ,  C07K14/00 ,  C07K17/00

CPC分类号： G01N33/5041 ,  A01K2217/05 ,  C07K14/705 ,  C07K2319/00 ,  C12Q1/6876 ,  C12Q2600/124 ,  C12Q2600/158 ,  G01N33/5008 ,  G01N33/566 ,  G01N2333/4719 ,  G01N2333/705 ,  G01N2333/726 ,  G01N2500/04

摘要： Heteromeric taste receptors are provided. These receptors comprise a first polypeptide containing extracellular domains and transmembrane domains wherein the extracellular domains are at least 95% identical to the extracellular domains of specific T1R1 polypeptides and the transmembrane domains are at least 95% identical to the corresponding transmembrane domains of the specific T1R1 polypeptide or a different GPCR; and a second polypeptide comprising extracellular and transmembrane domains wherein the extracellular domains are at least 95% identical to the extracellular domains of specific T1R3 polypeptides and the transmembrane domains are at least 95% identical to the corresponding transmembrane domains of the specific T1R3 polypeptide or a different GPCR.

摘要翻译： 提供异味味觉受体。 这些受体包含含有细胞外结构域和跨膜结构域的第一多肽，其中细胞外结构域与特异性T1R1多肽的细胞外结构域至少95％相同，并且跨膜结构域与特异性T1R1多肽的相应跨膜结构域至少95％相同 或不同的GPCR; 和包含细胞外和跨膜结构域的第二多肽，其中细胞外结构域与特异性T1R3多肽的细胞外结构域至少95％相同，并且跨膜结构域与特异性T1R3多肽的相应跨膜结构域至少95％相同或不同 GPCR。

公开(公告)号：US08071719B2

公开(公告)日：2011-12-06

申请号：US12270834

申请日：2008-11-13

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： C07K1/00 ,  C07K14/00 ,  C07K17/00

CPC分类号： G01N33/5041 ,  A01K2217/05 ,  C07K14/705 ,  C07K2319/00 ,  C12Q1/6876 ,  C12Q2600/124 ,  C12Q2600/158 ,  G01N33/5008 ,  G01N33/566 ,  G01N2333/4719 ,  G01N2333/705 ,  G01N2333/726 ,  G01N2500/04

摘要： Heteromeric taste receptors are provided. These receptors comprise a first polypeptide which comprises extracellular and transmembrane domains wherein the transmembrane domains are at least 95% identical to the transmembrane domains of specific T1R1 polypeptides, and the extracellular domains are at least 95% identical to the corresponding extracellular domains of the specific T1R1 polypeptide or a different GPCR; and a second polypeptide which comprises extracellular and transmembrane domains wherein the transmembrane domains are at least 95% identical to the transmembrane domains of specific T1R3 polypeptides, and the extracellular domains are at least 95% identical to the corresponding extracellular domains of the specific T1R3 polypeptide or that of a different GPCR.

摘要翻译： 提供异味味觉受体。 这些受体包含第一多肽，其包含细胞外和跨膜结构域，其中跨膜结构域与特异性T1R1多肽的跨膜结构域至少95％相同，并且细胞外结构域与特异性T1R1的相应胞外结构域至少95％相同 多肽或不同的GPCR; 和包含细胞外和跨膜结构域的第二多肽，其中所述跨膜结构域与特异性T1R3多肽的跨膜结构域至少95％相同，并且细胞外结构域与特异性T1R3多肽的相应细胞外结构域至少95％相同，或 不同的GPCR。

公开(公告)号：US07927825B2

公开(公告)日：2011-04-19

申请号：US12270830

申请日：2008-11-13

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： C01N33/567 ,  C07K1/00 ,  C07K14/00 ,  C07K17/00

CPC分类号： G01N33/5041 ,  A01K2217/05 ,  C07K14/705 ,  C07K2319/00 ,  C12Q1/6876 ,  C12Q2600/124 ,  C12Q2600/158 ,  G01N33/5008 ,  G01N33/566 ,  G01N2333/4719 ,  G01N2333/705 ,  G01N2333/726 ,  G01N2500/04

摘要： Methods of quantifying the taste of compounds for food and beverages are provided. These methods comprise contacting the compounds with an isolated heteromeric receptor comprising at least one T1R2 polypeptide and at least one T1R3 polypeptide.

摘要翻译： 提供了定量食品和饮料化合物味道的方法。 这些方法包括使化合物与包含至少一种T1R2多肽和至少一种T1R3多肽的分离的异聚受体接触。

公开(公告)号：US20090221798A1

公开(公告)日：2009-09-03

申请号：US12270857

申请日：2008-11-14

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： C07K14/00

CPC分类号： G01N33/5008 ,  A01K2217/05 ,  C07K14/705 ,  G01N33/5041 ,  G01N33/566 ,  G01N2333/726

摘要： Heteromeric taste receptors are provided. These receptors comprise a first polypeptide which comprises extracellular and transmembrane domains wherein the transmembrane domains are at least 95% identical to the transmembrane domains of specific T1R2 polypeptides, and the extracellular domains are at least 95% identical to the corresponding extracellular domains of the specific T1R2 polypeptide or a different GPCR; and a second polypeptide which comprises extracellular and transmembrane domains wherein the transmembrane domains are at least 95% identical to the transmembrane domains of specific T1R3 polypeptides, and the extracellular domains are at least 95% identical to the corresponding extracellular domains of the specific T1R3 polypeptide or that of a different GPCR.

摘要翻译： 提供异味味觉受体。 这些受体包含第一多肽，其包含细胞外和跨膜结构域，其中跨膜结构域与特异性T1R2多肽的跨膜结构域至少95％相同，并且细胞外结构域与特异性T1R2的相应胞外结构域至少95％相同 多肽或不同的GPCR; 和包含细胞外和跨膜结构域的第二多肽，其中所述跨膜结构域与特异性T1R3多肽的跨膜结构域至少95％相同，并且细胞外结构域与特异性T1R3多肽的相应细胞外结构域至少95％相同，或 不同的GPCR。

公开(公告)号：US20090221001A1

公开(公告)日：2009-09-03

申请号：US12270838

申请日：2008-11-13

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： G01N33/566

CPC分类号： G01N33/5008 ,  C12N2503/00 ,  C12N2510/00 ,  C40B30/04 ,  G01N33/6872 ,  G01N2333/726 ,  G01N2500/10

摘要： Methods for identifying compounds that modulate the T1R1/T1R3 umami taste receptors are provided. These methods comprise screening for compounds that compete with lactisole for binding to and/or inhibiting the T1R1/T1R3 umami taste receptor.

摘要翻译： 提供了鉴定调节T1R1 / T1R3鲜味受体的化合物的方法。 这些方法包括筛选与乳清蛋白竞争结合和/或抑制T1R1 / T1R3鲜味受体的化合物。

公开(公告)号：US20080262087A1

公开(公告)日：2008-10-23

申请号：US11932949

申请日：2007-10-31

申请人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

发明人： Mark Zoller ,  Xiaodong Li ,  Lena Staszewski ,  Shawn O'Connell ,  Sergey Zozulya ,  Jon Elliot Adler ,  Hong Xu ,  Fernando Echeverri

IPC分类号： A61K31/192 ,  C07K14/705 ,  C12N5/08 ,  C12N5/06 ,  C07K17/00 ,  A61P27/00 ,  G01N33/566 ,  G01N33/53 ,  C40B30/04 ,  C12Q1/68

CPC分类号： G01N33/5041 ,  A01K2217/05 ,  C07K14/705 ,  C07K2319/00 ,  C12Q1/6876 ,  C12Q2600/124 ,  C12Q2600/158 ,  G01N33/5008 ,  G01N33/566 ,  G01N2333/4719 ,  G01N2333/705 ,  G01N2333/726 ,  G01N2500/04

摘要： The present invention relates to the discovery that the T1R receptors assemble to form functional taste receptors. Particularly, it has been discovered that co-expression of T1R1 and T1R3 results in a taste receptor that responds to umami taste stimuli, including monosodium glutamate. Also, it has been discovered that co-expression of the T1R2 and T1R3 receptors results in a taste receptor that responds to sweet taste stimuli including naturally occurring and artificial sweeteners.Also the present invention relates to the use of hetero-oligomeric taste receptors comprising T1R1/T1R3 and T1R2/T1R3 in assays to identify compounds that respectively respond to umami taste stimuli and sweet taste stimuli.Further, the invention relates to the constitutive of cell lines that stably or transiently co-express a combination of T1R1 and T1R3; or T1R2 and T1R3; under constitutive or inducible conditions. The use of these cells lines in cell-based assays to identify umami and sweet taste modulatory compounds is also provided, particularly high throughput screening assays that detect receptor activity by use of fluorometric imaging.Finally, the invention relates to the discovery that some compounds, e.g., lactisole, inhibit both the activities of human T1R2/T1R3 and T1R1/T1R3 receptors, and accordingly the sweet and umami taste, suggesting that these receptors may be the only sweet and umami receptors.