Methods for identifying compounds that modulate mammalian tub protein activity

• US 5,861,239 A
• Filed: 09/02/1997
• Issued: 01/19/1999
• Est. Priority Date: 04/12/1996
• Status: Expired due to Fees

• Current Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Original Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Inventors: Kleyn, Patrick W., Moore, Karen J., Kapeller, Rosana
• Primary Examiner(s): RAILEY, JOHNNY F
• Application Number:
  US08/922,267
• Time in Patent Office: 504 Days
• Field of Search: 435/4
• US Class Current: 435/4
• 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: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: C07K 14/5759 Products of obesity genes, e.g. leptin, obese (OB), tub, fat

Abstract

The present invention relates to the identification of novel nucleic acid molecules and proteins encoded by such nucleic acid molecules or degenerate variants thereof, that participate in the control of mammalian body weight. The nucleic acid molecules of the present invention represent the gene corresponding to the mammalian tub gene, a gene that is involved in the regulation of body weight. The present invention also relates to methods for identifying compounds that modulate tub protein activity.

Claims

1. A method for identifying compounds that modulate tub protein activity, comprising:

(a) contacting a cell expressing a tub protein, an extract thereof, or an isolated tub protein, in the presence or absence of a test compound with a SH2 containing peptide,

(b) determining whether the test compound alters the interaction of the tub protein with the SH2 containing peptide, and

(c) selecting the test compound that alters the interaction of the tub protein with the SH2 containing peptide

wherein the tub protein comprises;

(i) the amino acid sequence shown in SEQ ID NO;

2,

(ii) amino acid residues 1-133 shown in SEQ ID NO;

2,

(iii) amino acid residues 190-505 shown in SEQ ID NO;

2,

(iv) amino acid residues 1-133 and 190-505 shown in SEQ ID NO;

2,

(v) the amino acid sequence shown in SEQ ID NO;

8,

(vi) amino acid residues 1-133 shown in SEQ ID NO;

8,

(vii) amino acid residues 190-506 shown in SEQ ID NO;

8,

(viii) amino acid residues 1-133 and 190-506 shown in SEQ ID NO;

8,

(ix) an amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69856,

(x) the amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 97222,

(xi) the amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 97221,

(xii) the amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69874,

(xiii) the amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69857,

(xiv) an amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69858, or

(xv) an amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69859.

2. The method of claim 1 in which the SH2 containing peptide is selected from the group comprising PLC gamma, Abl, Lck, Hck, Fgr, BLk, Src, Fyn, Yes and Lyn kinase.

3. The method of claim 1 in which at least one tyrosine residue of the tub protein is phosphorylated.

4. The method of claim 1 in which the tub protein is contained in an isolated cellular extract.

5. The method of claim 1 which the tub protein is expressed recombinantly.

6. The method of claim 1 in which the tub protein is phosphorylated.

7. A method for identifying compounds that modulate tub protein activity, comprising:

(a) contacting a cell expressing a tub protein, an extract thereof, or an isolated tub protein, in the presence or absence of a test compound with a SH2 containing peptide,

(b) determining whether the test compound alters the interaction of the tub protein with the SH2 containing peptide, and

(c) selecting the test compound that alters the interaction of the tub protein with the SH2 containing peptide,

wherein the tub protein is encoded by an allele of a nucleotide sequence encoding;

(i) the amino acid sequence shown in SEQ ID NO;

2,

(ii) amino acid residues 1-133 shown in SEQ ID NO;

2,

(iii) amino acid residues 190-505 shown in SEQ ID NO;

2,

(iv) amino acid residues 1-133 and 190-505 shown in SEQ ID NO;

2,

(v) the amino acid sequence shown in SEQ ID NO;

8,

(vi) amino acid residues 1-133 shown in SEQ ID NO;

8,

(vii) amino acid residues 190-506 shown in SEQ ID NO;

8,

(viii) amino acid residues 1-133 and 190-506 shown in SEQ ID NO;

8,

(ix) an amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69856,

(x) the amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 97222,

(xi) the amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 97221,

(xii) the amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69874,

(xiii) the amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69857,

(xiv) an amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69858, or

(xv) an amino acid sequence encoded by the nucleic acid insert of the clone contained in ATCC Deposit No. 69859,

and wherein said allele is encoded by a tub gene locus and said nucleotide sequence hybridizes under stringent wash condition, 0.1×

SSC, 0.1% SDS at 65°

C., to the complement of a nucleotide sequence encoding any of (i)-(xv).

8. The method of claim 7 in which the SH2 containing peptide is selected from the group comprising PLC gamma, Abl, Lck, Hck, Fgr, BLk, Src, Fyn, Yes, and Lyn kinase.

9. The method of claim 7 in which at least one tyrosine residue of the tub protein is phosphorylated.

10. The method of claim 7 in which the tub protein is contained in an isolated cellular extract.

11. The method of claim 7 in which the tub protein is expressed recombinantly.

12. The method of claim 7 in which the tub protein is phosphorylated.