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重组肝细胞生长因子激活剂抑制剂1:在果蝇S2细胞中的表达、纯化及结晶

Recombinant hepatocyte growth factor activator inhibitor 1: expression in Drosophila S2 cells, purification and crystallization.

作者信息

Liu Min, Yuan Cai, Jiang Yunbin, Jiang Longguang, Huang Mingdong

机构信息

State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Science, Fuzhou, Fujian 350002, People's Republic of China.

出版信息

Acta Crystallogr F Struct Biol Commun. 2017 Jan 1;73(Pt 1):45-50. doi: 10.1107/S2053230X16020082.

Abstract

Hepatocyte growth factor activator inhibitor 1 (HAI-1) is a multi-domain membrane-associated protease inhibitor that potently inhibits a variety of serine proteases such as hepatocyte growth factor activator and matriptase. Different truncates of HAI-1 show varying potencies for inhibition of target proteases, suggesting that the domain organization of HAI-1 plays a critical role in its function. Here, the soluble full-length extracellular part of HAI-1 (sHAI-1) was expressed using the Drosophila S2 insect-cell expression system. Diffraction-quality crystals of sHAI-1 were produced using ammonium sulfate as precipitant. The crystal diffracted to 3.8 Å resolution and belonged to space group P422, with unit-cell parameters a = b = 95.42, c = 124.50 Å. The asymmetric unit contains one sHAI-1 molecule.

摘要

肝细胞生长因子激活剂抑制剂1(HAI-1)是一种多结构域膜相关蛋白酶抑制剂,能有效抑制多种丝氨酸蛋白酶,如肝细胞生长因子激活剂和matriptase。HAI-1的不同截短形式对靶蛋白酶的抑制效力不同,这表明HAI-1的结构域组织在其功能中起关键作用。在此,利用果蝇S2昆虫细胞表达系统表达了HAI-1的可溶性全长细胞外部分(sHAI-1)。以硫酸铵作为沉淀剂获得了sHAI-1的衍射质量晶体。该晶体的衍射分辨率为3.8 Å,属于空间群P422,晶胞参数a = b = 95.42,c = 124.50 Å。不对称单元包含一个sHAI-1分子。

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