25934, a novel fatty acid desaturase and uses therefor

• US 6,686,185 B1
• Filed: 11/28/2000
• Issued: 02/03/2004
• Est. Priority Date: 03/07/2000
• Status: Expired due to Fees

• Current Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Original Assignee: Millennium Pharmaceuticals Incorporated (Takeda Pharmaceutical Company Limited)
• Inventors: Logan, Thomas Joseph, Glucksmann, Maria Alexandra
• Primary Examiner(s): Slobodyansky, Elizabeth
• Application Number:
  US09/723,806
• Time in Patent Office: 1,162 Days
• Field of Search: 435/189, 435/6, 435/252.3, 435/320.1, 435/325, 536/23.2, 536/23.5
• US Class Current: 435/189
• 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: C12N 9/0071 acting on paired donors with incorporation of molecular oxygen (1.14)
• CPC Class Codes: C12N 9/0083 Miscellaneous (1.14.99)

Abstract

The invention provides isolated nucleic acids molecules, designated 25934 nucleic acid molecules, which encode a novel desaturase family member. The invention also provides antisense nucleic acid molecules, recombinant expression vectors containing 25934 nucleic acid molecules, host cells into which the expression vectors have been introduced, and nonhuman transgenic animals in which a 25934 gene has been introduced or disrupted. The invention still further provides isolated 25934 proteins, fusion proteins, antigenic peptides and anti-25934 antibodies. Diagnostic methods utilizing compositions of the invention are also provided. The invention also provides methods for modulating fatty acid metabolism utilizing the compositions of the invention. Accordingly, methods of treating, preventing and/or diagnosing cardiovascular disorders, such atherosclerosis, hypertriglyceridemia, hypercholesterolemia, and hyperlipidemia, are disclosed.

Claims

1. An isolated nucleic acid comprising a nucleotide sequence that encodes a polypeptide comprising the sequence of SEQ ID NO:

2.

2. The nucleic acid of claim 1, wherein the polypeptide consists of the amino acid sequence of SEQ ID NO:

2.

3. An isolated nucleic acid comprising the nucleotide sequence of SEQ ID NO:

3 or a complete complement thereof.

4. The nucleic acid of claim 3, wherein the nucleic acid consists of the nucleotide sequence of SEQ ID NO:

3.

5. The nucleic acid of claim 3, wherein the nucleic acid comprises the nucleotide sequence of SEQ ID NO:

1 or a complete complement thereof.

6. An isolated nucleic acid comprising a nucleotide sequence that encodes a polypeptide comprising an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO:

2 or the sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number 2167, wherein the percent identity is determined using the ALIGN program in the GCG software package, using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4, and wherein the polypeptide has fatty acid desaturase activity.

7. The nucleic acid of claim 6, wherein the amino acid sequence is at least 90% identical to the sequence of SEQ ID NO:

2 or the sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number 2167.

8. The nucleic acid of claim 7, wherein the amino acid sequence is at least 95% identical to the sequence of SEQ ID NO:

2 or the sequence encoded by the cDNA insert of the plasmid deposited with the ATCC as Accession Number 2167.

9. The nucleic acid of claim 8, wherein the polypeptide comprises amino acid residues 51-295 of SEQ ID NO:

2.

10. The nucleic acid of claim 6, further comprising a sequence encoding a heterologous polypeptide.

11. A vector comprising the nucleic acid of claim 6.

12. The vector of claim 11, wherein the vector comprises nucleic acid sequences which regulate expression of a polypeptide encoded by the nucleic acid.

13. A host cell comprising the vector of claim 12.

14. The host cell of claim 13, which is a mammalian host cell.

15. A method for producing a polypeptide, the method comprising culturing the host cell of claim 14 under conditions in which the nucleic acid is expressed to produce the polypeptide.

16. An isolated nucleic acid comprising a nucleotide sequence that is at least 80% identical to the nucleotide sequence of SEQ ID NO:

3, wherein the percent identity is determined using the NBLAST program with a score of 100 and a word length of 12, and wherein the nucleotide sequence encodes a polypeptide having fatty acid desaturase activity.

17. The nucleic acid of claim 16, wherein the nucleotide sequence is at least 90% identical to the nucleotide sequence of SEQ ID NO:

3.

18. The nucleic acid of claim 17, wherein the nucleotide sequence is at least 95% identical to the nucleotide sequence of SEQ ID NO:

3.

19. The nucleic acid of claim 16, wherein the polypeptide comprises amino acid residues 51-295 of SEQ ID NO:

2.

20. An isolated nucleic acid of at least 750 nucleotides in length comprising a nucleotide sequence that hybridizes to the sequence of SEQ ID NO:

3 or the complete complement thereof under conditions of hybridization at 45°

C. in 6.0×

sodium chloride/sodium citrate (SSC) followed by one or more washes in 0.2×

SSC, 0.1% SDS at 65°

C., wherein the nucleotide sequence encodes a polypeptide having fatty acid desaturase activity.

21. The nucleic acid of claim 20 wherein the nucleic acid comprises at least 800 nucleotides.

22. The nucleic acid of claim 21, wherein the nucleic acid comprises at least 1000 nucleotides.

23. The nucleic acid of claim 22, wherein the nucleic acid comprises at least 1200 nucleotides.

24. The nucleic acid of claim 23, wherein the nucleic acid comprises at least 1400 nucleotides.