切割后的硫氧还蛋白融合蛋白能够使难溶性雌激素受体α（ERα）与合成配体形成的复合物结晶。
Cleaved thioredoxin fusion protein enables the crystallization of poorly soluble ERalpha in complex with synthetic ligands.
作者信息
Cura Vincent, Gangloff Monique, Eiler Sylvia, Moras Dino, Ruff Marc
机构信息
Laboratoire de Biologie et Génomique Structurales, IGBMC, Illkirch, France.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Jan 1;64(Pt 1):54-7. doi: 10.1107/S1744309107066444. Epub 2007 Dec 20.
Abstract
The ligand-binding domain (LBD) of human oestrogen receptor alpha was produced in Escherichia coli as a cleavable thioredoxin (Trx) fusion in order to improve solubility. Crystallization trials with either cleaved and purified LBD or with the purified fusion protein both failed to produce crystals. In another attempt, Trx was not removed from the LBD after endoproteolytic cleavage and its presence promoted nucleation and subsequent crystal growth, which allowed the structure determination of two different LBD-ligand-coactivator peptide complexes at 2.3 A resolution. This technique is likely to be applicable to other low-solubility proteins.
摘要
为提高溶解性，人雌激素受体α的配体结合结构域（LBD）在大肠杆菌中作为可裂解的硫氧还蛋白（Trx）融合蛋白产生。对裂解并纯化的LBD或纯化的融合蛋白进行结晶试验均未能产生晶体。在另一次尝试中，内蛋白水解切割后未从LBD中去除Trx，其存在促进了成核和随后的晶体生长，从而得以在2.3埃分辨率下确定两种不同的LBD-配体-共激活剂肽复合物的结构。该技术可能适用于其他低溶解性蛋白。
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