NS4 nucleic acids and polypeptides and methods of use for the treatment of body weight disorders

An isolated nucleic acid molecule comprising DNA which comprises at least about 80% sequence identity to (a) a DNA molecule encoding the same mature polypeptide encoded by the human cDNA deposited with the ATCC on (i) May 15, 2001 under ATCC Deposit No. PTA-3376 (DNA146649-1789R1), (ii) Apr. 4, 2000 under ATCC Deposit No. PTA-1627 or (iii) May 31, 2000 under ATCC Deposit No. PTA-1971 (DNA149995-2871), or (b) the complement of the DNA molecule of (a).

An isolated nucleic acid molecule comprising DNA having at least about 80% nucleic acid sequence identity to the full-length polypeptide coding sequence of the human cDNA deposited with the ATTC on:

• (a) (i) May 15, 2001 under ATCC Deposit No. PTA-3376 (DNA146649-1789R1), (ii) Apr. 4, 2000 under ATCC Deposit No. PTA-1627 or (iii) May 31, 2000 under ATCC Deposit No. PTA-1971 (DNA149995-2871);
• or (b) the complement of the DNA molecule of (a).

An isolated nucleic acid molecule encoding a NS4 polypeptide comprising DNA that hybridizes under at least moderately stringent conditions to the complement of the nucleic acid sequence that encodes amino acids (i) 1 or about 20 to about 87, inclusive, of FIG. 2A (SEQ ID NO:4);

• (ii) 1 or about 20 to about 95, inclusive, of FIG. 2B (SEQ ID NO:5) or (iii) 1 or about 20 or about 95, inclusive, of FIG. 2C (SEQ ID NO:6), wherein the hybridization occurs in the presence of 20% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6). 5× Denhardt's solution, 10% dextran sulfate, and 20 μg/ml denatured sheared salmon sperm DNA, followed by washing the filters in 1×SSC at about 37° C.-50° C.

An isolated nucleic acid molecule comprising at least 31 nucleotides and which is produced by hybridizing a test DNA molecule under stringent hybridization conditions with (a) a DNA molecule which encodes a NS4 polypeptide comprising a sequence of amino acid residues from about (i) 1 or about 20 to about 87, inclusive, of FIG. 2A (SEQ ID NO:4);

• (ii) 1 or about 20 to about 95, inclusive, of FIG. 2B (SEQ ID NO:5) or (iii) 1 or about 20 or about 95, inclusive, of FIG. 2C (SEQ ID NO:6), or (b) the complement of the DNA molecule of (a), and isolating the test DNA molecule.

A nucleic acid molecule deposited with the ATCC under accession number (i) PTA-3376 (DNA146649-1789R1), (ii) PTA-1627 or (iii) PTA-1971 (DNA149995-2871).

A process for producing a NS4 polypeptide comprising culturing the host said NS4 polypeptide from the cell culture.

An isolated NS4 polypeptide comprising an amino acid sequence comprising at least about 80% sequence identity to the sequence of amino acid residues from about (i) 1 or about 20 to about 87, inclusive, of FIG. 2A (SEQ ID NO:4);

• (ii) 1 or about 20 to about 95, inclusive, of FIG. 2B (SEQ ID NO:5) or (iii) 1 or about 20 or about 95, inclusive, of FIG. 2C (SEQ ID NO:6).

An isolated NS4 polypeptide having at least about 80% sequence identity to the polypeptide encoded by the cDNA insert of the vector deposited with the ATCC on (i) May 15, 2001 under ATCC Deposit No. PTA-3376 (DNA146649-1789R1), (ii) Apr. 4, 2000 under ATCC Deposit No. PTA-1627 or (iii) May 31, 2000 under ATCC Deposit No. PTA-1971 (DNA149995-2871).

An isolated NS4 polypeptide comprising the sequence of amino acid residues from (i) 1 or about 20 to about 87 of FIG. 2A (SEQ ID NO:4);

• (ii) 1 or about 20 to about 95, inclusive, of FIG. 2B (SEQ ID NO:5) or (iii) 1 or about 20 or about 95, inclusive of FIG. 2C (SEQ ID NO:6), or a fragment thereof sufficient to provide a binding site for an anti-NS4 antibody.

An isolated polypeptide produced by (i) hybridizing a test DNA molecule under stringent conditions with (a) a DNA molecule encoding a NS4 polypeptide comprising the sequence of amino acid residues from (i) 1 or about 20 to about 87, inclusive, of FIG. 2A (SEQ ID NO:4);

• (ii) 1 or about 20 to about 95, inclusive, of FIG. 2B (SEQ ID NO:5) or (iii) 1 or about 20 or about 95, inclusive, of FIG. 2C (SEQ ID NO:6), or (b) the complement of the DNA molecule of (a), (ii) culturing a host cell comprising said test DNA molecule under conditions suitable for the expression of said polypeptide, and (iii) recovering said polypeptide from the cell culture.

A chimeric molecule comprising an NS4 polypeptide fused to a heterologous amino acid sequence.

An antibody which specifically binds to an NS4 polypeptide.

A composition of matter comprising (a) an NS4 polypeptide, (b) an agonist to a NS4 polypeptide, (c) an antagonist to a NS4 polypeptide, or (d) an anti-NS4 antibody in admixture with a pharmaceutically acceptable carrier.

A method for screening for a bioactive agent capable of binding to NS4 comprising:

• a) adding a candidate bioactive agent to a sample of NS4;
• and
• b) determining the binding of said candidate agent to said NS4, wherein binding indicates a bioactive agent capable of binding to NS4.

A method for screening for a bioactive agent capable of modulating the activity of NS4, said method comprising the steps of:

• a) adding a candidate bioactive agent to a sample of NS4;
• and
• b) determining an alteration in the biological activity of NS4, wherein an alteration indicates a bioactive agent capable of modulating the activity of NS4.

A method of identifying a receptor for NS4, said method comprising combining NS4 with a composition comprising cell membrane material wherein said NS4 complexes with a receptor on said cell membrane material, and identifying said receptor as a NS4 receptor.

A method of treating an individual for a body weight disorder, said method comprising administering to said individual a composition comprising a therapeutically effective amount of NS4.

A method of reducing total body mass in an individual, said method comprising administering to said individual a therapeutically effective amount of NS4.

A method of reducing the level of at least one triglyceride or free fatty acid in an individual, said method comprising administering to said individual a therapeutically effective amount of NS4.

The method according to claim 60, wherein said NS4 has at least about 80% amino acid sequence identity to the amino acid sequence shown in FIG. 2A (SEQ ID NO:4), FIG. 2B (SEQ ID NO:5) or FIG. 2C (SEQ ID NO:6).

A method of increasing the metabolic rate in an individual, said method comprising administering to said individual an effective amount of NS4.

The method according to claim 65, wherein said NS4 has at least about 80% amino acid sequence identity to the amino acid sequence shown in FIG. 2A (SEQ ID NO:4), FIG. 2B (SEQ ID NO:5) or FIG. 2C (SEQ ID NO:6).

A rodent comprising a genome comprising a transgene encoding NS4.