人组织蛋白酶D的纯化与结晶

Purification and crystallization of human cathepsin D.

作者信息

Fusek M, Baudys M, Metcalf P

机构信息

European Molecular Biology Laboratory, Heidelberg, Germany.

出版信息

J Mol Biol. 1992 Jul 20;226(2):555-7. doi: 10.1016/0022-2836(92)90968-p.

DOI:10.1016/0022-2836(92)90968-p

PMID:1640466

Abstract

The two-chain form of human cathepsin D was purified from human spleen with a method utilizing an ion exchange chromatography step prior to the pepstatin affinity column normally used to purify aspartic proteases. The protein was crystallized from 21% polyethylene glycol 8000 at pH 4.0 using the hanging drop vapour diffusion method. Small crystals were used as seeds to grow crystals suitable for X-ray data collection. The crystals diffract to a resolution of 3.2 A and have space group P2(1)2(1)2(1) with unit cell dimensions a = 59.9 A, b = 99.6 A, c = 133.6 A. There are two molecules in the asymmetric unit.

摘要

人组织蛋白酶D的双链形式是从人脾脏中纯化得到的，其方法是在通常用于纯化天冬氨酸蛋白酶的胃蛋白酶抑制剂亲和柱之前，利用离子交换色谱步骤。使用悬滴气相扩散法，该蛋白质在pH 4.0的21%聚乙二醇8000中结晶。小晶体用作种子来生长适合X射线数据收集的晶体。这些晶体的衍射分辨率为3.2 Å，空间群为P2(1)2(1)2(1)，晶胞尺寸a = 59.9 Å，b = 99.6 Å，c = 133.6 Å。不对称单位中有两个分子。