Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...
Patent
1996-05-16
2000-05-23
Chambers, Jasemine C.
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
Introduction of a polynucleotide molecule into or...
435440, 435 691, 435183, 536 231, 536 235, C12N 1509, C12N 900, C12P 2106, C07H 2104
Patent
active
060665025
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to endothelin converting enzymes, to processes for their preparation and to their use.
Elevated endothelin levels are thought to be associated with a large number of disorders such as essential hypertension, myocardial infarct, acute kidney failure, shock or heart failure. An association of this type is also suggested by the results obtainable with endothelin antibodies. In animal models it was possible therewith to reduce the size of a myocardial infarct dose-dependently (Watanabe et al., Nature 344, 114 (1990)), to have a beneficial effect on kidney function (Kon et al., J. Clin. Invest. 83, 1762 (1989)), and to reduce the nephrotoxicity of cyclosporin (Kon et al., Kidney Int. 37, 1487 (1990)).
Endothelin converting enzyme (ECE-1) liberates endothelin 1 from the precursor molecule big endothelin 1 which consists of 38 amino acids. The unwanted biological effects caused by endothelin 1 can be counteracted, for example, by inhibiting endothelin converting enzyme and thus endothelin 1 biosynthesis.
Enzyme activities which liberate endothelins or endothelin-like molecules from the corresponding precursor molecules, the big endothelins, have been isolated from various cell lines (Takeda et al., Biochem. Biophys. Res. Comm. 176, 860 (1991), Ohnaka et al., Biochem. Biophys. Res. Comm. 168, 1128 (1990); Matsumura et al. FEBS Lett. 293, 45 (1992), Ahn et al. Proc. Natl. Acad. Sci. U.S.A. 89, 8606 (1992), PCT WO 92/13944, Ohnaka et al. Clin. Exp. Hypertension A 14; No. 4 (1992), Okada et al. Biochem. Biophys. Res. Comm. 180, 1019 (1991), Takada et al. Biochem. Biophys. Res. Comm. 182, 1383 (1992), Opgenorth et. al. FASEB 6, 2653 (1992)).
However, these enzyme activities have not to date been adequately characterized. They are in the form of enzyme mixtures which have not been greatly concentrated. However, impure enzyme mixtures of these types are not very suitable for use in assay methods for finding specific ECE inhibitors because foreign proteases which are of no physiological relevance considerably interfere with the enzyme assay.
Thus, big endothelin is cleaved, for example, also by the serine protease chymothrypsin [sic] as well as by papain, thermolysin and NEP 24.11 to endothelin and endothelin-like products which are incorrectly assigned to the enzyme ECE in assay methods. As a consequence, there are contradictions in the description of endothelin converting enzyme in the literature.
The statements about the molecular weight of endothelin converting enzyme vary from 65 KDalton to 540 KDalton; the statements about Km and vmax values likewise differ considerably from one another (Opgenorth et al. FASEB 6, 2653 (1992); Sawamura et al. Biochem. Biophys. Acta 1161,295 (1993)). There are also contradictions about whether endothelin converting enzyme is to be assigned to the family of aspartate proteases or of metalloproteases (Sawamura et al. Biochem. Biophys. Acta 1161,295 (1993); Biochem. Pharmacol. 43,845 (1992). Furthermore, contradictory statements about characteristic information as to whether the metalloprotease is a cytoplasmic or membrane-bound enzyme, and which substrates are converted by the particular ECE (big Et-1; big Et-3), are to be found in the literature: Matsumura et al. (FEBS Lett. 293, 45 (1992)): Takahashi et al. (J. Biol. Chem. 268;21394 (1993)); Okada et al. (Biochem. Biophys. Res. Comm. 180, 1019 (1991)); Ohnaka et al. (Clin. Exp. Hypertension A 14; No. 4 (1992)); Matsumura et al. (FEBS Lett. 305;86 (1992)).
To date no unambiguous description of ECE making it possible satisfactorily to define ECE has been shown. Such a definition may be possible only through determination of the primary structure, of the amino acid sequence, of ECE. For this it is necessary first to prepare ECE in pure form.
It is an object of the present invention to prepare endothelin converting enzyme in pure form.
We have found that this object is achieved by an endothelin converting enzyme having a molecular weight of 250,000 Dalton and the partial amino acid sequence SEQ ID NO: 1.
The
REFERENCES:
Shipp et al. Molecular cloning of the common acute lymphoblastic leukemia antigen (CALLA) identifies a type II integral membrane protein. PNAS, vol. 85, pp. 4819-4823, Jul. 1988.
Opgenorth et al. Endothelian-converting enzymes. FASEB Journal, vol. 6, No. 9, pp. 2653-2659, Jun. 1992.
J. Of Bio. Chem., vol. 269, No. 28, Jul. 199, pp. 18275-18278, 1994.
J. of Bio. Chem., vol. 268, No. 35, Dec. 15, pp. 26759-26766, 1993.
Bio. and Biophysical Comm., vol. 203,. No. 3, 1994, pp. 1417-1422.
Pharmaceutical Res., vol. 5, No. 9, 1988.
Bio. and Biophysical Res. Comm., vol 171, No. 3, 1990, Sep. 28, 1990, pp. 1192-1198.
Bioorganic & Med. Chem. Lt. vol. 3, No. 10, 1993.
Bioscience, Biotech. and Biochem., vol. 57, Nov. 1993, pp. 1944-1945.
Derwent, JP 4 120 593, Abst.
Jor. of Bio. Chem. vol. 268, No. 28, Oct. 5, 1993, pp. 21394-21398.
Hillen Heinz
Jacob Elard
Kroger Burkhard
Meyer Thomas
Otter Rainer
BASF - Aktiengesellschaft
Chambers Jasemine C.
Clark Deborah J. R.
LandOfFree
Endothelin converting enzyme (ECE) does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Endothelin converting enzyme (ECE), we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Endothelin converting enzyme (ECE) will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1836178