Partial purification and characterization of a novel neutral proteinase from human uterine cervix

摘要: 1. Human uterine cervical stroma was found to contain a Ca2+-independent neutral proteinase against casein and N-benzoyl-dl-arginine p-nitroanilide (Bz-dl-Arg-Nan). This enzyme tightly bound an insoluble material (20000g pellet) solubilized by high concentrations of NaCl or KCl. High them in the reaction system, however, inhibited reversibly activity this enzyme. 2. The partially purified extraction with NaCl, gel filtration on Sephadex G-200 affinity chromatography casein–Sepharose. 3. optimal pH 7.4–8.0 Bz-dl-Arg-Nan. molecular weight be about 1.4×105 G-200. 4. significantly di-isopropyl phosphorofluoridate (0.1mm). concentration phenylmethanesulphonyl fluoride (5mm), 7-amino-1-chloro-3-l-tosylamidoheptan-2-one (0.5mm), antipain (10μm) leupeptin also inhibitory, but chymostatin (40μg/ml), soya-bean trypsin inhibitor (2.5mg/ml), human plasma (10%, v/v), p-chloromercuribenzoate (1mm), EDTA (10mm) 1-chloro-4-phenyl-3-l-tosylamidobutan-2-one (1mm) had no effect 5. hydrolysed casein, Bz-dl-Arg-Nan heat-denatured collagen, inactive towards native collagen several synthetic substrates, such as 4-phenylazobenzyloxycarbonyl-Pro-Leu-Gly-Pro-d-Arg, 3-carboxypropionyl-Ala-Ala-Ala 2,4-dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-d-Arg, proteoglycan. did not act plasminogen activator. 6. These properties suggested that cervix different from enzymes previously reported.