Partial purification and characterization of human sperminogen.
Partial purification and characterization of human sperminogen.
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人精蛋白原的部分纯化和表征。
DOI：
10.1095/biolreprod36.4.1063
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发表时间：
1987
影响因子：
3.6
通讯作者：
Polakoski,KL
中科院分区：
文献类型：
--
作者：
Siegel,MS;Bechtold,DS;Willand,JL;Polakoski,KL
最近从人类精子中纯化了一种被称为精子原的非顶体酶原，并对其一些性质进行了测定。纯化过程包括在pH 3.0条件下对洗涤的射精进行酸提取，然后用Sephadex G-75超细柱凝胶过滤。精原从柱子上洗脱成一条与顶体酶原完全分离的单一条带。明胶-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(明胶-十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法)证实了这种分离。这一酶谱还表明，最终的精原制剂含有四种形式的酶原，分子量在32,000到36,000之间。在中性的pH条件下，精子原转化为其酶活性形式的精蛋白，产生典型的酶原自激活的S形曲线。几个因素对这种自转率的影响表明，精子原和顶体酶原之间存在着特定的差异。精氨酸水解N-α-苯甲酰基-L-精氨酸乙酯(BzArgOEt)，并被利马豆胰酶抑制剂、胰腺胰酶抑制剂、N-乙酰基-L-亮氨酰-L-亮氨酰-L-精氨酸(亮胃素)和对甲苯磺酰-L-赖氨酸氯甲基酮所抑制，表明该酶具有胰酶样特异性，可能属于胰酶样酶类。由于顶体酶通常被认为是哺乳动物精子中唯一的类胰酶，因此，人类精蛋白原和精子蛋白的存在需要进一步探讨精子蛋白系统的功能和相互关系。
A recently recognized non-proacrosin zymogen referred to as sperminogen has been purified from human spermatozoa, and several of its properties have been determined. The purification procedure included acid extraction of washed ejaculated sperm at pH 3.0, followed by gel filtration of the solubilized extract over a Sephadex G-75 superfine column. The sperminogen eluted from the column in a single band that was completely separated from the proacrosin band. This separation was confirmed by a gelatin-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (gelatin-SDS-PAGE) zymograph. This zymograph also demonstrated that the final sperminogen preparation contained four forms of zymogen, with molecular weights between 32,000 and 36,000. At neutral pH, the sperminogen was converted into spermin, its enzymatically active form, yielding a sigmoidal curve typical of zymogen autoactivation. The effects of several factors on the rate of this autoconversion indicate specific differences between sperminogen and proacrosin. Spermin hydrolyzed N-α-benzoyl-L-arginine ethyl ester (BzArgOEt), and was inhibited by lima bean trypsin inhibitor, pancreatic trypsin inhibitor, N-acetyl-L-leucyl-L-leucyl-L-argininal (leupeptin), and tosyl-L-lysine chloromethyl ketone, indicating that the enzyme has a trypsin-like specificity and probably belongs to the class of trypsin-like enzymes. Since acrosin is generally believed to be the only trypsin-like enzyme in mammalian sperm, the demonstration of human sperminogen and spermin necessitates further inquiry into the functions and the relationships between sperm proteinase systems.