J Integr Plant Biol. ›› 2003, Vol. 45 ›› Issue (3): 289-294.

• Research Articles • Previous Articles     Next Articles

Growth of Large Single Crystals of Nitrogenase CrFe Protein and MnFe Protein

Lű Yu-Bing, ZHAO Ying, ZHAO Jian-Feng, DAI Xiao-Hu, CANG Huai-Xing, WANG Yao-Ping and HUANG Ju-Fu   


By using the liquid/liquid diffusion method at a suitable crystallization conditions, large single and dark brown crystals (the sides of the largest crystals were 0.20 mm×0.20 mm×0.07 mm and 0.18 mm×0.18 mm×0.05 mm, respectively) could be obtained from the solutions of nitrogenase CrFe protein and MnFe protein purified from a mutant UW3 of Azotobacter vinelandii Lipmann grown in Cr- or Mn-containing but NH3-free medium. The time of crystal formation, as well as the number,size, shape and quality of crystals obviously depended on the concentrations of PEG, MgCl2 and NaCl. The liquid/liquid diffusion method seems to benefit CrFe protein and MnFe protein for the growth of large single crystals for X-ray diffraction analysis. Key words: mutant UW3 of Azotobacter vinelandii; nitrogenase CrFe protein and MnFe protein; growth of large single crystals

吕玉兵  赵 颖  赵剑峰  代小虎  仓怀兴  王耀萍 黄巨富

(1. 中国科学院植物研究所光合作用与环境分子生理学重点实验室,北京100093;2. 中国科学院生物物理研究所,北京100101)

摘要: 用液/液扩散法,从分别含Cr和Mn的无氨培养基中生长的固氮菌(Azotobacter vinelandii Lipmann)突变种UW3中纯化出的CrFe蛋白和MnFe蛋白,在合适的结晶条件下生长出深棕色大单晶(最大晶体的尺寸分别为0.20mm×0.20mm×0.07mm和0.18mm×0.18mm×0.05mm)。PEG、MgCl2和NaCl的浓度对这两种蛋白的出晶时间、晶核数目、晶体大小和形状都有明显影响。结果表明,用此结晶法有利于CrFe蛋白和MnFe蛋白生长出可供X-射线衍射分析的大单晶。

关键词: 棕色固氮菌突变种UW3;固氮酶铬铁蛋白和锰铁蛋白;大单晶生长

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