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光系统I与其天然电子受体铁氧化还原蛋白复合物的结晶及电子顺磁共振表征

Crystallization and electron paramagnetic resonance characterization of the complex of photosystem I with its natural electron acceptor ferredoxin.

作者信息

Fromme Petra, Bottin Hervé, Krauss Norbert, Sétif Pierre

机构信息

Max Volmer Laboratorium, Institut für Chemie, Fakultät II, Technische Universität Berlin, Germany.

出版信息

Biophys J. 2002 Oct;83(4):1760-73. doi: 10.1016/S0006-3495(02)73942-8.

Abstract

The formation of a transient complex between photosystem I and ferredoxin is involved in the process of ferredoxin photoreduction in oxygenic photosynthetic organisms. Reduced ferredoxin is an essential redox intermediate involved in many assimilatory processes and is necessary for the reduction of NADP(+) to NADPH. Single crystals from a complex of photosystem I with ferredoxin were grown using PEG 400 and CaCl(2) as precipitation agents. The crystals diffract x-rays to a resolution of 7-8 A. The space group was determined to be orthorhombic with the unit cell dimensions a = 194 A, b = 208 A, and c = 354 A. The crystals contain photosystem I and ferredoxin in a 1:1 ratio. Electron paramagnetic resonance (EPR) measurements on these crystals are reported, where EPR signals of the three [4Fe-4S] clusters F(A), F(B), F(X), and the [2Fe-2S] cluster of ferredoxin were detected. From the EPR spectra observed at three particular orientations of the crystal in the magnetic field, the full orientation pattern of the F g-tensor was simulated. This simulation is consistent with the presence of 12 magnetically inequivalent F clusters per unit cell with the C(3) axis of the PSI trimers oriented at (23 degrees, 72 degrees, 77 degrees ) to the unit cell axes.

摘要

光系统I与铁氧还蛋白之间形成的瞬态复合物参与了光合生物中铁氧还蛋白光还原过程。还原态铁氧还蛋白是许多同化过程中必不可少的氧化还原中间体,也是将NADP(+)还原为NADPH所必需的。使用聚乙二醇400和氯化钙作为沉淀剂,培养了光系统I与铁氧还蛋白复合物的单晶。这些晶体对X射线的衍射分辨率为7-8埃。确定其空间群为正交晶系,晶胞尺寸a = 194埃,b = 208埃,c = 354埃。晶体中光系统I和铁氧还蛋白的比例为1:1。报道了对这些晶体的电子顺磁共振(EPR)测量,其中检测到了三个[4Fe-4S]簇F(A)、F(B)、F(X)以及铁氧还蛋白的[2Fe-2S]簇的EPR信号。根据在磁场中晶体三个特定取向观察到的EPR光谱,模拟了F g张量的完整取向模式。该模拟结果与每个晶胞中存在12个磁不等价的F簇一致,光系统I三聚体的C(3)轴与晶胞轴的夹角为(23度、72度、77度)。

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