title: Antipain and leupeptin restrict uterine DNA synthesis and function in mice.
abstract: As in rats, administration of estradiol to ovariectomized mice results in a trypsin-like proteolytic activity in the uterus. After fractionation of uteri from estradiol-treated ovariectomized mice the protease activity was found in the 12,000 times g pellet and the nucleus, appearing first in the former. Further fractionation of the pellet by discontinuous sucrose gradient centrigugation resulted in sedimentation of the protease with 5'-nucleotidase, a marker enzyme for plasma membrane and separate from mitochondrial and lysosomal enzyme markers. Solubilization was best accomplished by lysis at 37 degrees. The soluble enzyme from mouse uterus had optimal activity at about 43 degrees and pH 8.3 and was inhibited by diisopropylfluorophosphate, tosylarginine methyl ester, antipain, and leupeptin, but not by soybean trypsin inhibitor. Inhibition in vitro by antipain and leupeptin, two low molecular weight peptides, prompted the study of their effect in vivo on the mouse uterus. After intact, cycling female mice received subcutaneous injections of antipain and leupeptin for 16 days, their uteri showed significant diminution in weight and total DNA when compared to untreated controls. Fertility rates were also diminished. Trypsin-like protease activity may be essential to normal uterine metabolism and function.
mesh_terms: D000818:Animals; D002369:Castration; D004247:DNA / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D005298:Fertility / Q000187:drug effects; D007700:Kinetics; D007976:Leupeptins / Q000494:pharmacology*; D051379:Mice; D009842:Oligopeptides / Q000494:pharmacology*; D009929:Organ Size / Q000187:drug effects; D011480:Protease Inhibitors; D014599:Uterus / Q000187:drug effects / Q000378:metabolism*
chemical_list: D007976:Leupeptins; D009842:Oligopeptides; D011480:Protease Inhibitors; D004958:Estradiol; D004247:DNA
references.citation: Biochem J. 1960 Mar;74:450-6
references.pmid: 14444755
references.citation: Biochem J. 1956 Feb;62(2):315-23
references.pmid: 13293190
references.citation: Biochem J. 1953 Aug;55(1):170-1
references.pmid: 13093635
references.citation: Science. 1952 Nov 28;116(3022):591-4
references.pmid: 13015119
references.citation: J Biol Chem. 1951 Apr;189(2):665-70
references.pmid: 14832284
references.citation: Arch Biochem Biophys. 1976 Oct;176(2):411-6
references.pmid: 185962
references.citation: J Cell Biol. 1976 Nov;71(2):472-86
references.pmid: 993259
references.citation: J Biol Chem. 1976 Sep 25;251(18):5694-702
references.pmid: 965386
references.citation: Nature. 1970 Jul 11;227(5254):170-1
references.pmid: 5428405
references.citation: J Biol Chem. 1974 Jul 10;249(13):4295-305
references.pmid: 4368727
references.citation: J Exp Med. 1973 Jan 1;137(1):85-111
references.pmid: 4347290
references.citation: Biochem Biophys Res Commun. 1973 Nov 1;55(1):84-90
references.pmid: 4274574
references.citation: Arch Biochem Biophys. 1976 Mar;173(1):347-54
references.pmid: 4033
references.citation: J Cell Biol. 1975 Jan;64(1):182-210
references.pmid: 162790
references.citation: Endocr Res Commun. 1974;1(4):331-7
references.pmid: 4462994
references.citation: Biochemistry. 1975 Nov 4;14(22):4923-7
references.pmid: 1182129