Patent EA-003293-B1 (EA)
Title: Polypeptides encoded by a human lipase-like gene, compositions containing them and methods of use of compositions
Inventor: JAYE MICHAEL C
IPC: C12R1/91
Filed: 1997-12-05
Abstract: 1. An isolated polypeptide encoded by a lipase like gene, wherein the polypeptide (a) binds heparin, (b) has homology with human lipoprotein lipase and hepatic lipase, and (c) comprises a 39 kD catalytic domain of the triacylglycerol lipase family, wherein said polypeptide has an amino acid sequence selected from group consisting of SEQ ID NO: 6 and SEQ ID NO: 8 2. The isolated polypeptide of claim 1, wherein the 39 kD catalytic domain of the triacylglycerol lipase family has the amino acid sequence of SEQ ID NO: 10. 3. The isolated polypeptide of claim l, wherein the polypeptide has an amino acid sequence of SEQ ID NO: 8 and an apparent molecular weight of about 55 kD on a 10% SDS-PAGE gel. 4. The isolated polypeptide of claim 1, wherein the polypeptide has an amino acid sequence of SEQ ID NO: 8 and an apparent molecular weight of about 68 kD on a 10% SDS-PAGE gel. 5. A composition comprising the polypeptide of claim 1 and a biologically compatible solution. 6. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier. 7. An isolated nucleic acid, which encodes the polypeptide of claim 1, wherein isolated nucleic acid has nucleotide sequence selected from the group consisting of SEQ ID NO: 5, and SEQ ID NO: 7, SEQ ID NO: 9 and nucleotides from 252-1754 of SEQ ID NO: 7. 8. An isolated nucleic acid which hybridizes at high stringency to a nucleic acid having a sequence of SEQ ID NOS: 5 and 7, and wherein the said conditions of high stringency represent 0.1 SSC, 0.5% SDS under temperature of 68 degree C. 9. The isolated nucleic acid of claim 8, which comprises 20 nucleotides. 10. The isolated nucleic acid of claim 8, which is an antisense nucleic acid. 11. The isolated nucleic acid of claim 10 wherein the antisense nucleic acid sequence is operably linked to a gene expression regulatory region. 12. The isolated nucleic acid, which hybridizes at high stringency to a target, selected from the group consisting of nucleotides from 1036-1065 of SEQ ID NO: 5. 13. A composition comprising the nucleic acid of claim 10 and a biologically compatible solution. 14. A pharmaceutical composition comprising the nucleic acid of claim 10 and a pharmaceutically acceptable carrier. 15. A vector comprising the isolated nucleic acid of claims 7-8 operably linked to a regulatory region. 16. The vector of claim 15 wherein the regulatory region is from a heterologous source. 17. The vector of claims 15 or 16 which is a viral vector. 18. The vector of claim 17, which is an adenoviral vector. 19. Culture of eukaryotic cells, comprising vector by claims 15-18, wherein the said cells are selected from group, consisting of cells PsiCRIP, cells GP+envA-12, cells CHO, cells THP-1, cells HUVEC, HCEAC and cells COS-7. 20. Cell culture by claim 20, wherein the said cells are cells COS-7. 21. A composition comprising vector of claim 15-18 and a biologically compatible solution. 22. A pharmaceutical composition comprising the vector of claims 15-18 and a pharmaceutically acceptable carrier. 23. A method for preparing a polypeptide comprising culturing the recombinant cell according to claims 19-21 under conditions permitting the expression of the polypeptide, with the following recovery of polypeptide from recombinant cells. 24. A method of screening for agonists or antagonists of LLG activity comprising: (a) contacting potential agonists or antagonists with LLG and a substrate of LLG by claim 1, and (b) measuring the ability of the potential agonists or antagonists to enhance or inhibit LLG activity. 25. A method for the enzymatic hydrolysis of a phosphatidylcholine ester comprising contacting said phosphatidylcholine ester with the polypeptide of claim 1. 26. A method of improving the serum lipid profile of a human or other animal having an undesirable lipid profile comprising administration of an effective amount of the pharmaceutical composition of claim 6 or 14. 27. 1. An isolated polypeptide encoded by a lipase like gene, wherein the polypeptide (a) binds heparin, (b) has homology with human lipoprotein lipase and hepatic lipase, and (c) comprises a 39 kD catalytic domain of the triacylglycerol lipase family, (d) originates from rabbit, wherein said polypeptide has an amino acid sequence of SEQ ID NO: 12. 28. An isolated nucleic acid encoding the polypeptide by claims 27. 29. An isolated nucleic acid has a nucleotide sequence SEQ ID NO: 11. 30 A polyclonal antibody capable of specifically binding to the polypeptide of claim 1. 31. A polyclonal antibody neutralizing the phospholipase activity of the polypeptide of claim 1. 32. A composition comprising the polyclonal antibody of claims 30-31 and a biologically compatible solution. 33. A pharmaceutical composition comprising the polyclonal antibody of claims 30-31 and a pharmaceutically acceptable carrier.