Endopeptidase

Details Endopeptidase Endopeptidase

1993-10-06

1996-05-21

Wax, Robert A.

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

435 681, 435 691, 4351723, 536 232, 935 14, C12N 964, C12N 1557, C12N 1510, C07K 100

Patent

active

055189121

ABSTRACT:
There is provided by the invention a novel composition comprising, in at least partially purified form, an endopeptidase endogenous to biological cells. The endopeptidase is characterized by having a molecular weight of about 110,000 daltons as measured by SDS PAGE; by being substantially insensitive to PMSF, TPCK, E-64, leupeptin, bacitracin, phosphoramidon and pepstatin; by being substantially inhibited by EDTA and 1,10-phenanthroline; and by being capable of cleaving peptide substrates comprising an alpha helical structural of at least about twelve to fourteen amino acids wherein said helix has a hydrophobic face and a hydrophilic face, said cleavage occuring amino terminal to a lysine or arginine residue on the hydrophilic face positioned within the context of at least four nonpolar amino acids substantially aligned along the hydrophobic face of the helix.

REFERENCES:
Docherty, et al., "Proinsulin endopeptidase substrate specificities defined by site-directed mutagenesis of proinsulin," J. Biol. Chem. 264, 18335-18339 (1989).
Gomez et al.,"Site-specific mutagenesis identifies amino acid residues critical in prohormone processing," EMBO J. 8, 2911-2916 (1989).
Thorne, et al., "An in vivo characterization of the cleavage site specificity of the insulin cell prohormone processing enzymes," J. Biol. Chem. 265, 8436-8443 (1990).
Rholam, et al., "Precursors for peptide hormones share common secondary structures forming features at the proteolytic processing sites," FEBS Lett. 207, 1-6 (1986).
Bek, et al., "Prohormonal cleavage sites are associated with omega loops," Biochem. 29, 178-183 (1990).
von Heijne, et al., "Patterns of amino acids near signal-sequence cleavage sites," Eur. J. Biochem. 133, 17-21 (1983).
Duffaud, et al., "Signal peptidases recognize a structural feature at the cleavage site of secretory proteins," J. Biol. Chem. 263, 10224-10228 (1988).
Folz, et al., "Substrate specificity of eucaryotic signal peptidase," J. Biol. Chem. 263, 2070-2078 (1988).
Caulfield et al., "Synthetic substrate for eucaryotic signal peptidase," J. Biol. Chem. 264, 15813-15817 (1989).
Checler, et al., "Purification and characterization of novel neurotensin-degrading peptidase from rat brain synaptic membranes," J. Biol. Chem. 261, 11274-11281 (1986).
Clamagirand, et al., "Partial purification and functional properties of an endoprotease from bovine neurosecretory granules cleaving procytocin
europhysin peptides at the basic amino acid doublet," Biochem. 26, 6018-6023 (1987).
Gluschankof, et al., "Role of peptide substrate structure in the selective processing of peptide prohormones at basic amino acid pairs by endoproteases," FEBS Lett. 234, 149-152 (1988).
Loh, et al., "Proteolysis in neuropeptide processing and other neural functions," Annu. Rev. Neuro. 7, 189-222 (1984).
Beinfeld, et al., "Characterization of an endoprotease from rat small intestinal mucosal secretary granules which generates somatostatin-28 from prosomatostatin by cleavage after a single arginine residue," J. Biol. Chem. 264, 4460-4465 (1989).
Brakch, et al., "Processing endoprotease recognizes a structural feature at the cleavage site of peptide prohormones," J. Biol. Chem. 264, 15912-15916 (1989).
Hurt, et al., "Amino-terminal deletions in the presequence of an imported mitocondrial protein block the targeting function and proteolytic cleavage of the presequence at the carboxy terminus," J. Biol. Chem. 262, 1420-1424 (1987).
Zasloff, et al., "Antimicrobial activity of synthetic magainin peptides and several analogues," Proc. Natl. Acad. Sci. USA 85, 910-913 (1988).
Bevins, et al., "Peptides from frog skin," Annu. Rev. Biochem. 59, 395-414 (1990).
Giovannini, et al., "Biosynthesis and degradation of peptides derived from Xenopus laevis prohormones," Biochem J. 243, 113-120 (1987).
Richter, et al., "Sequence of preprocaerulein cDNAs cloned from skin of Xenpus laevis," J. Biol. Chem. 261, 3676-3680 (1986).
Poulter, et al., "Levitide, a neurohormone-like peptide from the skin of Xenopus laevis," J. Biol. Chem. 263, 3279-3283 (1988).
Gibson, et al., "Novel peptide fragments originating from PGLa and the caerulein and xenopsin precursors from Xenopus laevis," J. Biol. Chem. 261, 5341-5349 (1986).
Deutscher, M. (Ed.) "Methods in Enzymology" vol. 182 pp. 9-15, 285-421, 459-477, 602-613, 738-751 pub. 1990 by Acad. Press Inc.
Kullmann, W. "Enzymatic Peptide Synthesis" pub. 1987 by CRC Press, Inc. pages-Preface, 37,-40, 61-82.
Kimmel et al. "Methods in Enzymology" vol. 152 pub. 1987 by Acad. Press Inc. pp. 661-704.
Belyavsky, A. et al. "PCR based cDNA Library Construction . . . " Nuc. Acids. Res. vol. 17 pp. 2919-2932 (Apr. 1989).
Deutscher, M. (ed.) Meth. in Enzymology vol. 182 pp. 9-15, 285-421, 459-477, 603-613, 738-751 (1990).
Kimmel et al. Meth. in Enzymology vol. 152 pp. 661-704 (1987).
Belyansky et al. Nuc. Acids Res. 17:2919-2932 (1989).

Affiliated with

Also associated with

Rating

Endopeptidase does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Endopeptidase, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Endopeptidase will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2037334