人酸性α-D-甘露糖苷酶的纯化及化学特性分析

Purification and chemical characterization of human acidic alpha-D-mannosidase.

作者信息

Lee J E, Little T E, Yoshida A

出版信息

Enzyme. 1982;28(1):33-40. doi: 10.1159/000459082.

Abstract

The acidic alpha-D-mannosidase (EC 3.2.1.24) has been purified from human placentae. Milligram quantities of the enzyme were obtained from several placentae, using a step-wise purification procedure which includes Con A-Sepharose treatment, acetone precipitation, heat treatment, DEAE-cellulose column chromatography and preparative disc electrophoresis. A high degree of purity of the purified enzyme was demonstrated by acrylamide gel electrophoresis, isoelectric focusing, and sedimentation equilibrium centrifugation. Immunological homogeneity of the preparation was demonstrated by a single precipitin line between the antiserum and purified, or partially purified enzyme preparation. The amino acid and carbohydrate composition of the enzyme was determined. The enzyme was found to be a glycoprotein containing 13.5% carbohydrate. The molecular weight of the enzyme was estimated at 205,000 +/- 18,400.

摘要

酸性α-D-甘露糖苷酶（EC 3.2.1.24）已从人胎盘中纯化出来。通过逐步纯化程序，从多个胎盘中获得了毫克量的该酶，该程序包括伴刀豆球蛋白A-琼脂糖处理、丙酮沉淀、热处理、DEAE-纤维素柱色谱和制备性圆盘电泳。通过丙烯酰胺凝胶电泳、等电聚焦和沉降平衡离心证明了纯化酶的高纯度。抗血清与纯化的或部分纯化的酶制剂之间的单一沉淀线证明了该制剂的免疫同质性。测定了该酶的氨基酸和碳水化合物组成。发现该酶是一种含糖量为13.5%的糖蛋白。该酶的分子量估计为205,000±18,400。