Goldbaum F A, Polikarpov I, Cauerhff A A, Velikovsky C A, Braden B C, Poljak R J
Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Buenos Aires, Argentina.
J Struct Biol. 1998 Oct;123(2):175-8. doi: 10.1006/jsbi.1998.4022.
Lumazine synthase from Brucella abortus was overexpressed in Escherichia coli, refolded, and purified to apparent homogeneity. Crystals of lumazine synthase were grown by the hanging drop vapor diffusion method using polyethylene glycol 8000 or ammonium sulfate as precipitants. They belong to the trigonal space group P321 with cell parameters a = b = 132.00A, c = 167.25 A. A complete diffraction data set to 3.7 A resolution has been collected using synchrotron radiation. Preliminary analysis of the quaternary structure of this protein by means of a self-rotation function calculated with the diffraction data clearly indicates 532 symmetry compatible with the presence of an icosahedral lumazine synthase particle.
来自流产布鲁氏菌的核黄素合酶在大肠杆菌中过量表达、复性并纯化至表观均一。使用聚乙二醇8000或硫酸铵作为沉淀剂,通过悬滴气相扩散法培养核黄素合酶晶体。它们属于三方晶系空间群P321,晶胞参数a = b = 132.00Å,c = 167.25 Å。利用同步辐射收集了分辨率为3.7 Å的完整衍射数据集。用衍射数据计算的自旋转函数对该蛋白质的四级结构进行初步分析,清楚地表明532对称与二十面体核黄素合酶颗粒的存在相符。