从牛多形核中性粒细胞中部分纯化碱性磷酸酶及其某些特性

Partial purification of alkaline phosphatase from bovine polymorphonuclear neutrophils and some properties.

作者信息

Yasuura S, Nagaoka I, Yamashita T, Namihisa T

出版信息

Comp Biochem Physiol B. 1985;82(4):587-93. doi: 10.1016/0305-0491(85)90494-8.

Abstract

Alkaline phosphatase was purified from bovine polymorphonuclear neutrophils by butanol extraction and a combination of ion exchange, gel filtration and affinity chromatography. The enzyme was partially purified 2300-fold with a 4.7% yield and a sp. act. of 206 units/mg of protein. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate indicated a single activity band with the mol. wt of 165,000. The pH optima for the enzyme were 10.0 with p-nitrophenylphosphate and phenylphosphate and were 9.0 when beta-glycerophosphate, AMP and ADP were used. The enzyme was activated by Mg2+, Mn2+, Co2+ and Ni2+ but was inhibited by Zn2+. The enzyme was inhibited by EDTA and the EDTA-inactivated enzyme was reactivated by Mg2+, Mn2+ and Co2+ but not Zn2+.

摘要

通过丁醇萃取以及离子交换、凝胶过滤和亲和层析相结合的方法，从牛多形核嗜中性粒细胞中纯化出碱性磷酸酶。该酶经过部分纯化，纯化倍数达2300倍，产率为4.7%，比活性为206单位/毫克蛋白质。十二烷基硫酸钠聚丙烯酰胺凝胶电泳显示有一条单一活性带，分子量为165,000。该酶作用于对硝基苯磷酸酯和苯磷酸酯时的最适pH为10.0，而作用于β-甘油磷酸酯、AMP和ADP时的最适pH为9.0。该酶被Mg2+、Mn2+、Co2+和Ni2+激活，但被Zn2+抑制。该酶被EDTA抑制，且被EDTA失活的酶可被Mg2+、Mn2+和Co2+重新激活，但不能被Zn2+重新激活。