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US 6,645,736 B2Granted
Calcium independent cytosolic phospholipase A2/B enzymes
AssigneeGenetics Institute, Inc.
InventorsSimon Jones, Jin Tang
Life of this patent
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Abstract
The invention provides a novel calcium-independent cytosolic phospholipase A2/B enzyme, polynucleotides encoding such enzyme and methods for screening unknown compounds for anti-inflammatory activity mediated by the arachidonic acid cascade.
Claims15
A method for identifying an inhibitor of phospholipase activity, said method comprising:
- a) contacting a phospholipid with a candidate inhibitor compound and a phospholipase enzyme peptide, wherein the phospholipase enzyme peptide comprises an amino acid sequence selected from:
- i) the amino acid sequence of SEQ ID NO:17;
- ii) the amino acid sequence of SEQ ID NO:19;
- iii) a fragment of the amino acid sequence of SEQ ID NO:19 having activity in a mixed micelle assay with 1-palmitoyl-2-[14 C]-arachidonyl-phosphatidylcholine;
- iv) the amino acid sequence of SEQ ID NO:21;
- v) a fragment of the amino acid sequence of SEQ ID NO:21 having activity in a mixed micelle assay with 1-palmitoyl-2-[14C]-arachidonyl-phosphatidylcholine;
- vi) the amino acid sequence of SEQ ID NO:23;
- and
- vii) a fragment of the amino acid sequence of SEQ ID NO:23 having activity in a mixed micelle assay with 1-palmitoyl-2-[14C]-arachidonyl-phosphatidylcholine;
- b) determining whether the phospholipase enzyme peptide activity is inhibited by the candidate inhibitor compound, thereby identifying an inhibitor of phospholipase activity.
7 more elements
A method for identifying an inhibitor of phospholipase activity, said method comprising:
- a) contacting a phospholipid with a candidate inhibitor compound and a phospholipase enzyme peptide, wherein the phospholipase enzyme peptide comprises a polypeptide selected from:
- i) a polypeptide encoded by a polynucleotide, wherein the polypeptide has activity in a mixed micelle assay with 1-palmitoyl-2-[14C]-arachidonyl-phosphatidylcholine, and wherein the polynucleotide hybridizes at 65° C. in 4×SSC to the complement of the nucleic acid sequence of SEQ ID NO:
- 18;
- ii) a polypeptide encoded by a polynucleotide, wherein the polypeptide has activity in a mixed micelle assay with 1-palmitoyl-2-[14C]-arachidonyl-phosphatidylcholine, and wherein the polynucleotide hybridizes at 65° C. in 4×SSC to the complement of the nucleic acid sequence of SEQ ID NO:
- 20;
- and
- iii) a polypeptide encoded by a polynucleotide, wherein the polypeptide has activity in a mixed micelle assay with 1-palmitoyl-2-[14C]-arachidonyl-phosphatidylcholine, and wherein the polynucleotide hybridizes at 65° C. in 4×SSC to the complement of the nucleic acid sequence of SEQ ID NO:
- 22;
- b) determining whether the phospholipase enzyme peptide activity is inhibited by the candidate inhibitor compound, thereby identifying an inhibitor of phospholipase activity.
6 more elements
Dependencies are inferred from the claim text.