Anderson R A, Beyler S A, Mack S R, Zaneveld L J
Biochem J. 1981 Nov 1;199(2):307-16. doi: 10.1042/bj1990307.
A high-molecular-weight form of acrosin (alpha-acrosin, EC 3.4.21.10) was extracted from spermatozoa obtained from frozen semen and purified over 300-fold. Purification was effected by sequential use of Sephadex G-150, CM-cellulose and DEAE-cellulose chromatography. Properties of human acrosin were compared with those of human pancreatic trypsin. The molecular weight (Mr) of acrosin (70000) was greater than that of trypsin (Mr 21000). Isoelectric points for acrosin (pI = 9.0) and trypsin (pI = 8.2) were also different. alpha-N-Benzoyl-L-arginine ethyl ester was hydrolysed 50% more rapidly by acrosin than by trypsin. Acrosin had similar kcat. values for the hydrolysis of esters with different acylating groups (i.e. benzoyl-L-arginine and p-tosyl-L-arginine esters). In contrast, trypsin had dissimilar kcat. values for the hydrolysis of esters with different acylating groups. Kinetic data argue against deacylation as the rate-limiting step in ester hydrolysis by acrosin. Acrosin was less sensitive than trypsin to inhibition by 7-amino-1-chloro-3-L-tosylamidoheptan-2-one ('TLCK'), di-isopropyl fluorophosphate and soya-bean trypsin inhibitor. D-Fructose and D-arabinose inhibited acrosin, but had no effect on trypsin. The data indicate that definite differences exist between human acrosin and trypsin.
从冷冻精液中获取的精子中提取出一种高分子量形式的顶体蛋白酶（α-顶体蛋白酶，EC 3.4.21.10），并将其纯化了300多倍。通过依次使用葡聚糖凝胶G - 150、CM - 纤维素和DEAE - 纤维素色谱法进行纯化。将人顶体蛋白酶的特性与人类胰蛋白酶的特性进行了比较。顶体蛋白酶的分子量（Mr 70000）大于胰蛋白酶的分子量（Mr 21000）。顶体蛋白酶（pI = 9.0）和胰蛋白酶（pI = 8.2）的等电点也不同。α - N - 苯甲酰 - L - 精氨酸乙酯被顶体蛋白酶水解的速度比被胰蛋白酶水解的速度快50%。顶体蛋白酶对具有不同酰化基团的酯（即苯甲酰 - L - 精氨酸酯和对甲苯磺酰 - L - 精氨酸酯）的水解具有相似的kcat值。相比之下，胰蛋白酶对具有不同酰化基团的酯的水解具有不同的kcat值。动力学数据表明，脱酰作用不是顶体蛋白酶催化酯水解的限速步骤。顶体蛋白酶比胰蛋白酶对7 - 氨基 - 1 - 氯 - 3 - L - 甲苯磺酰胺基庚烷 - 2 - 酮（“TLCK”）、二异丙基氟磷酸酯和大豆胰蛋白酶抑制剂的抑制作用更不敏感。D - 果糖和D - 阿拉伯糖抑制顶体蛋白酶，但对胰蛋白酶没有影响。数据表明，人顶体蛋白酶和胰蛋白酶之间存在明显差异。
