大鼠肾脏碱性磷酸酶的纯化与特性研究

Purification and characterization of alkaline phosphatase from rat kidney.

作者信息

Nose K

出版信息

J Biochem. 1976 Feb;79(2):283-8. doi: 10.1093/oxfordjournals.jbchem.a131069.

DOI:10.1093/oxfordjournals.jbchem.a131069

Abstract

Alkaline phosphatase [EC 3.1.3.1.] was purified about 250-fold from rat kidney, and its enzymological properties were studied. Kidney homogenate was extracted with n-butanol, passed through Sephadex G-200 and chromatographed on a DEAE-cellulose column. The peak from the DEAE-cellulose column was subjected to isoelectric focusing, and the alkaline phosphatase activity was separated into two peaks. The molecular weights of alkaline phosphatase in these peaks were 4.8.X10(4) and 1.0X10(5), as determined by SDS-polyacrylamide gel electrophoresis. Anti-serum against alkaline phosphatase from rat kidney was prepared, and was shown to neutralize the activity from kidney, liver or bone, but not that from intestine.

摘要

从大鼠肾脏中纯化出碱性磷酸酶[EC 3.1.3.1.]，纯化倍数约为250倍，并对其酶学性质进行了研究。肾脏匀浆用正丁醇提取，经葡聚糖凝胶G - 200过滤，再在二乙氨基乙基纤维素柱上进行层析。二乙氨基乙基纤维素柱的洗脱峰进行等电聚焦，碱性磷酸酶活性被分离为两个峰。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳测定，这些峰中碱性磷酸酶的分子量分别为4.8×10⁴和1.0×10⁵。制备了抗大鼠肾脏碱性磷酸酶的抗血清，结果表明该抗血清能中和来自肾脏、肝脏或骨骼的碱性磷酸酶活性，但不能中和来自肠道的碱性磷酸酶活性。