来自大鼠睾丸的镁离子激活酸性磷酸酶的纯化及某些性质

Purification and some properties of a Mg(2+)-activated acid phosphatase from rat testis.

Panara F, Angiolillo A, Di Rosa I, Fagotti A, Contenti S, Simoncelli F, Lorvik S, Pascolini R

Istituto di Anatomia Comparata, Università degli Studi di Perugia, Italy.

Int J Biochem. 1994 Jul;26(7):885-90. doi: 10.1016/0020-711x(94)90081-7.

Abstract

The acid phosphatase (AcPase, EC 3.1.3.2) IV from rat testicular tissue was purified to apparent homogeneity. 2. The enzyme displays a native molecular weight of 70 kDa determined on gel permeation chromatography on a Sephadex G-100 column and 68 kDa using linear 5-20% sucrose density gradient centrifugation. The subunit molecular weight on SDS-PAGE analysis is 67 kDa, suggesting that the enzyme is a monomeric protein. 3. The enzyme does not bind to Concanavaline A-Sepharose 4B column, indicating that it is not a glycoprotein. 4. The rat testis AcPase IV is a metal activated enzyme in which Mg2+ is the metal activating agent with a Ka = 0.88 x 10(-3) M. The Michaelis constant for p-nitrophenylphosphate, in the presence of saturating concentrations of Mg2+ ions, is 0.23 x 10(-3) M. 5. The enzyme preferentially hydrolyzes p-nitrophenylphosphate, phenylphosphate and ATP.

摘要

大鼠睾丸组织中的酸性磷酸酶（AcPase，EC 3.1.3.2）IV被纯化至表观均一。2. 该酶在Sephadex G - 100柱上进行凝胶渗透色谱测定的天然分子量为70 kDa，使用线性5 - 20%蔗糖密度梯度离心法测定为68 kDa。SDS - PAGE分析的亚基分子量为67 kDa，表明该酶是一种单体蛋白。3. 该酶不与伴刀豆球蛋白A - Sepharose 4B柱结合，表明它不是糖蛋白。4. 大鼠睾丸AcPase IV是一种金属激活酶，其中Mg2 +是金属激活剂，Ka = 0.88×10（-3）M。在Mg2 +离子饱和浓度存在下，对硝基苯磷酸酯的米氏常数为0.23×10（-3）M。5. 该酶优先水解对硝基苯磷酸酯、苯磷酸酯和ATP。