ISOLATED BITTER TASTE RECEPTOR POLYPEPTIDES

• US 20120015432A1
• Filed: 08/31/2011
• Published: 01/19/2012
• Est. Priority Date: 04/07/2000
• Status: Active Grant

• Current Assignee: Firmenich SA
• Original Assignee: Senomyx Inc
• Inventors: Adler, Jon Elliot
• Granted Patent: US 8,450,472 B2
• Time in Patent Office: Days
• US Class Current: 435/325
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: C07H 21/04 with deoxyribosyl as saccharide radical
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: G01N 2333/726 G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 2500/04 Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)
• CPC Class Codes: G01N 33/566 using specific carrier or receptor proteins as ligand binding reagents where possible specific carrier or receptor proteins are classified with their target compounds
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

The claimed invention relates to the discovery of a specific human taste receptor in the T2R taste receptor family, hT2R61 that responds to particular bitter compounds The present invention further relates to the use of this receptor in assays for identifying ligands that modulate the activation of this taste receptor. These compounds may be used as additives and/or removed from foods, beverages and medicinals in order to modify (block) T2R-associated bitter taste. A preferred embodiment is the use of the identified compounds as additives in foods, beverages and medicinals for blocking bitter taste.

Claims

1-137. -137. (canceled)

138. An isolated nucleic acid molecule selected from the group consisting of:

(i) an isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS;

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, or a fragment thereof which comprises at least 75 nucleotides;

(ii) an isolated cDNA or an insoluble RNA transcribed therefrom that encodes a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NOS;

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, and 24, or a fragment thereof which encodes at least 25 contiguous amino acids of said polypeptide;

(iii) an isolated nucleic acid molecule having at least 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOS;

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, or to a fragment thereof which comprises at least 100 contiguous nucleotides thereof and an isolated nucleic acid molecule comprising a nucleotide sequence selected from the group consisting of SEQ ID NO'"'"'S;

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, and (iv) an isolated nucleic acid molecule comprising a fragment of a nucleic acid selected from the group consisting of SEQ ID NOS;

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, wherein said fragment encodes the transmembrane or extracellular region of the polypeptide encoded by said nucleic acid selected from the group consisting of SEQ ID NOS;

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23.

139. The isolated nucleic acid molecule of claim 138 which is selected from the group consisting of SEQ ID NOS:

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, or a fragment thereof which comprises at least 75 contiguous nucleotides thereof.

140. The isolated nucleic acid molecule of claim 138 which encodes a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NOS:

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, and 24, or a fragment thereof encoding at least 25 contiguous amino acid residues of said polypeptide.

141. An isolated nucleic acid molecule according to claim 138 having at least 90% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID) NOS:

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, or a fragment thereof comprising at least 100 contiguous nucleotides of any of said sequences.

142. An isolated nucleic acid molecule according to claim 138 having at least 95% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOS:

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, or a fragment thereof comprising at least 100 contiguous nucleotides of any of said sequences.

143. An isolated nucleic acid molecule according to claim 138 having at least 96% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOS:

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, or a fragment thereof comprising at least 100 contiguous nucleotides of any of said sequences.

144. An isolated nucleic acid molecule according to claim 138 having at least 97% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOS:

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, or a fragment thereof comprising at least 100 contiguous nucleotides of any of said sequences.

145. An isolated nucleic acid molecule according to claim 138 having at least 98% sequence identity with a nucleic acid sequence selected from the group consisting of SEQ ID NOS:

1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 23, or a fragment thereof comprising at least 100 contiguous nucleotides of any of said sequences.

146. An isolated nucleic acid molecule according to claim 138 which encodes a polypeptide having at least 99% sequence identity with a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NOS:

2, 4, 6, 8, 10, 12, 14, 16, 18, 20, and 24, or a fragment thereof that comprises at least 40 contiguous amino acids thereof.

147. An isolated nucleic acid molecule according to claim 138 which encodes a polypeptide having 100% sequence identity with a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NOS:

2, 4, 6, 8, 16, 12, 14, 16, 18, 20, and 24, or a fragment thereof that comprises at least 40 contiguous amino acids thereof.

148. An isolated nucleic acid molecule according to claim 138 which is operably linked to a constitutive or regulatable promoter or which is directly or indirectly attached to a nucleic acid sequence encoding a chaperone protein or a fragment thereof.

149. An expression vector that comprises a nucleic acid sequence according claim 138.

150. The expression vector of claim 149 which is a mammalian, yeast, bacterial or insect expression vector.

151. A cell which is transfected or transformed with at least one nucleic acid sequence according to claim 138.

152. A cell according to claim 151, wherein the cell is mammalian.