Purification and properties of a human seminal proteinase.
Purification and properties of a human seminal proteinase.
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人精液蛋白酶的纯化和特性。
DOI：
10.1042/bj1261135
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发表时间：
1972
期刊：
影响因子：
--
通讯作者：
K. Moghissi
中科院分区：
文献类型：
--
作者：
F. Syner;K. Moghissi
1.本发明描述了一种纯化蛋白酶的方法，该蛋白酶存在于人精浆中，并且先前显示出在体外加速精子通过宫颈粘液的迁移。一个25倍的纯化，实现了在三个步骤，包括硫酸铵分级分离，色谱上的CM-纤维素和凝胶过滤。2.该酶具有与胰凝乳蛋白酶相似的性质：最适pH 7.5-8.0，底物选择性为酪蛋白、血红蛋白和苯甲酰酪氨酸乙酯，而不选择苯甲酰精氨酸乙酯; 33000.然而，它不受1 mm-二异丙基磷氟化物或1 mm金属阳离子的影响，在这方面与糜蛋白酶不同。3.该酶的性质与Lundquist等人（1955）在精浆中发现的胰凝乳蛋白酶样酶的性质非常相似。4.使用二甲基酪蛋白允许在比普通酪蛋白（10 mg/ml）高5倍的底物浓度（高达50 mg/ml）下进行酶测定。
1. A method is described for the purification of a proteinase, present in human seminal plasma and previously shown to accelerate migration of spermatozoa through cervical mucus in vitro. A 25-fold purification was achieved in three steps, consisting of ammonium sulphate fractionation, chromatography on CM-cellulose and gel filtration. 2. The enzyme displays some properties similar to chymotrypsin: pH optimum 7.5-8.0; substrate preference of casein, haemoglobin and benzoyltyrosine ethyl ester but not benzoylarginine ethyl ester; mol.wt. 33000. However, it is unaffected by 1mm-di-isopropyl phosphofluoridate or 1mm metal cations, and in this respect differs from chymotrypsin. 3. The properties of the enzyme strongly resemble those of the ;chymotrypsin-like' enzyme discovered in seminal plasma by Lundquist et al. (1955). 4. The use of dimethyl-casein permitted the performance of enzyme assays at substrate concentrations five times higher (up to 50mg/ml) than could be achieved with ordinary casein (10mg/ml).