Purification and some properties of a Mg2+-activated acid phosphatase from rat testis

International Journal of Biochemistry Pub Date : 1994-07-01 DOI:10.1016/0020-711X(94)90081-7

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

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引用次数: 3

Abstract

1.
1. The acid phosphatase (AcPase, EC 3.1.3.2) IV from rat testicular tissue was purified to apparent homogeneity.
2.
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.
3. The enzyme does not bind to Concanavaline A-Sepharose 4B column, indicating that it is not a glycoprotein.
4.
4. The rat testis AcPase IV is a metal activated enzyme in which Mg²⁺ is the metal activating agent with a K_a, = 0.88 × 10⁻³ M. The Michaelis constant for p-nitrophenylphosphate, in the presence of saturating concentrations of Mg²⁺ ions, is 0.23 × 10⁻³ M.
5.
5. The enzyme preferentially hydrolizes p-nitrophenylphosphate, phenylphosphate and ATP.

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Mg2+活化大鼠睾丸酸性磷酸酶的纯化及性能研究

1.1. 从大鼠睾丸组织中纯化出酸性磷酸酶(AcPase, EC 3.1.3.2) IV，具有明显的同质性。该酶在Sephadex G-100柱上凝胶渗透色谱测定的天然分子量为70 kDa，在线性5-20%蔗糖密度梯度离心测定的天然分子量为68 kDa。SDS-PAGE分析亚基分子量为67 kDa，表明该酶为单体蛋白。该酶不与Concanavaline a - sepharose 4B柱结合，表明它不是糖蛋白。大鼠睾丸AcPase IV是一种金属活化酶，其中Mg2+为金属活化剂，Ka = 0.88 × 10−3 m。在Mg2+离子饱和浓度下，对硝基苯基磷酸的Michaelis常数为0.23 × 10−3 M.5.5。该酶优先水解对硝基苯基磷酸、苯基磷酸和ATP。

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International Journal of Biochemistry

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