Alternatively spliced isoform of acetyl-CoA carboxylase 2 (ACC2)

• US 7,150,969 B2
• Filed: 06/03/2005
• Issued: 12/19/2006
• Est. Priority Date: 06/04/2004
• Status: Expired due to Fees

• Current Assignee: Rosetta Inpharmatics LLC (Merck & Co., Inc.)
• Original Assignee: Rosetta Inpharmatics LLC (Merck & Co., Inc.)
• Inventors: Garrett-Engele, Philip W., Raymond, Christopher K., Kan, Zhengyan, Armour, Christopher D., Castle, John C.
• Primary Examiner(s): Prouty, Rebecca E.
• Assistant Examiner(s): Chowdhury, Iqbal
• Application Number:
  US11/144,368
• Publication Number:
  US 20050272082A1
• Time in Patent Office: 564 Days
• Field of Search: 435/183, 435/23.2, 435/6
• US Class Current: 435/6.18
• CPC Class Codes: C12N 9/93 Ligases (6)

Abstract

The present invention features nucleic acids and polypeptides encoding novel splice variant isoforms of acetyl-CoA carboxylase 2 (ACC2). The polynucleotide sequence of ACC2sv1 is provided by SEQ ID NO 3. The amino acid sequence of ACC2sv1 is provided by SEQ ID NO 4. The present invention also provides methods for using ACC2sv1 polynucleotides and proteins to screen for compounds that bind to ACC2sv1.

Claims

1. A method of screening for a compound that modulates activity of acetyl coenzyme A carboxylase 2, splice variant 1 (ACC2sv1) comprising:

(a) expressing a recombinant nucleic acid encoding ACC2sv1 comprising SEQ ID NO 4 in a cell;

(b) contacting said cell or a cell extract thereof with a test preparation comprising one or more test compounds; and

(c) measuring the effect of said test preparation on enzyme activity.

2. The method of claim 1, wherein said recombinant nucleic acid encoding ACC2sv1 also comprises an expression vector.

3. The method of claim 1, wherein said test preparation contains a compound that is an ACC inhibitor.

4. The method of claim 1, wherein said cell expressing a recombinant nucleic acid encoding ACC2sv1 comprising SEQ ID NO 4 is homogenized to obtain a cell extract and step (b) is performed by contacting said cell extract with said test preparation.

5. The method of claim 1, wherein said enzyme activity is measured using a CO₂ fixation assay.

6. The method of claim 1 wherein said measuring is performed using a high throughput format.

7. The method of claim 1 wherein, said test preparation comprises one or more agonists, antagonists, or allosteric modulators of ACC2sv1.

8. The method of claim 4 additionally comprising comparing said measured level of ACC2sv1 activity to the level of acetyl coenzyme A carboxylase activity in a control cell not expressing a recombinant nucleic acid encoding ACC2sv1.

9. The method of claim 4 additionally comprising comparing said measured level of ACC2sv1 activity to the level of acetyl coenzyme A carboxylase (ACC) activity in a cell expressing a recombinant nucleic acid encoding an ACC isofform polypeptide that is not ACC2sv1.