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Expression purification and crystallization of Streptococcus dysgalactiae-derived mitogen

机译:链球菌dysgalactiae衍生的有丝分裂原的表达纯化和结晶

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摘要

Superantigens are bacterial or viral toxins with potent immunostimulatory properties. Streptococcus dysgalactiae-derived mitogen, a 25 kDa protein, is a recently discovered superantigen isolated from S. dysgalactiae culture supernatant. Sequence considerations suggest that it belongs to a new superantigen family distinct from other superantigens. The protein was expressed in Escherichia coli cells and purified to homogeneity. Crystals were grown at pH 4.2–4.4 in the presence of 18–20%(w/v) PEG 3350 and 0.4 M lithium nitrate. A complete data set to 2.4 Å resolution was collected from a single crystal at liquid-nitrogen temperatures using synchrotron radiation. The crystals belong to space group P3/P31/P32, with unit-cell parameters a = b = 52.7, c = 62.4 Å, γ = 120° and one molecule in the crystallographic asymmetric unit.
机译:超抗原是具有有效免疫刺激特性的细菌或病毒毒素。链球菌痢疾小乳头来源的促分裂原是一种25 proteinkDa的蛋白质,是最近发现的从痢疾志贺菌培养物上清液中分离出来的超抗原。序列方面的考虑表明它属于不同于其他超抗原的新超抗原家族。该蛋白质在大肠杆菌细胞中表达,并纯化至同质。在18-20%(w / v)PEG 3350和0.4μM硝酸锂的存在下,晶体在pH 4.2-4.4的条件下生长。使用同步加速器辐射,在液氮温度下,从单晶收集了完整的数据,分辨率达到2.4Å。晶体属于P3 / P31 / P32空间群,晶胞参数a = b = 52.7,c = 62.4,γ= 120°,晶体学不对称单元中有一个分子。

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