Crystallization and preliminary crystallographic characterization of the extrinsic PsbP protein of photosystem II from Spinacia oleracea.
Crystallization and preliminary crystallographic characterization of the extrinsic PsbP protein of photosystem II from Spinacia oleracea.
复制标题
DOI：
10.1107/s1744309108040578
复制
发表时间：
2009-02
期刊：
影响因子：
--
通讯作者：
Jaroslava Kohoutová;I. Smatanova;J. Brynda;M. Lapkouski;J. Revuelta;J. B. Arellano;R. Ettrich
中科院分区：
文献类型：
--
作者：
Jaroslava Kohoutová;I. Smatanova;J. Brynda;M. Lapkouski;J. Revuelta;J. B. Arellano;R. Ettrich
利用N端His标记的重组PSBP蛋白在大肠杆菌中高效表达，对菠菜光系统II外源PSBP蛋白进行了初步的X射线衍射分析。以聚乙二醇550 MME为沉淀剂，硫酸锌为添加剂，采用静置蒸气扩散法，在pH 6.00的双Tris缓冲液中对重组PSBP蛋白(凝血酶消化的重组His标记PSBP)进行结晶。对溶解的晶体进行了SDS-PAGE分析，结果表明晶体中不含有重组PSBP蛋白的降解产物。PSBP晶体在P2(1)2(1)2(1)空间群中的衍射率为2.06 A，晶胞参数a=38.68，b=46.73，c=88.9A。
Preliminary X-ray diffraction analysis of the extrinsic PsbP protein of photosystem II from spinach (Spinacia oleracea) was performed using N-terminally His-tagged recombinant PsbP protein overexpressed in Escherichia coli. Recombinant PsbP protein (thrombin-digested recombinant His-tagged PsbP) stored in bis-Tris buffer pH 6.00 was crystallized using the sitting-drop vapour-diffusion technique with PEG 550 MME as a precipitant and zinc sulfate as an additive. SDS-PAGE analysis of a dissolved crystal showed that the crystals did not contain the degradation products of recombinant PsbP protein. PsbP crystals diffracted to 2.06 A resolution in space group P2(1)2(1)2(1), with unit-cell parameters a = 38.68, b = 46.73, c = 88.9 A.