Department of Biophysical Chemistry, Groningen Biological Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
J Biol Chem. 2011 Dec 30;286(52):44878-87. doi: 10.1074/jbc.M111.301101. Epub 2011 Nov 2.
In this work we have purified the Photosystem I (PSI) complex of Chlamydomonas reinhardtii to homogeneity. Biochemical, proteomic, spectroscopic, and structural analyses reveal the main properties of this PSI-LHCI supercomplex. The data show that the largest purified complex is composed of one core complex and nine Lhca antennas and that it contains all Lhca gene products. A projection map at 15 Å resolution obtained by electron microscopy reveals that the Lhcas are organized on one side of the core in a double half-ring arrangement, in contrast with previous suggestions. A series of stable disassembled PSI-LHCI intermediates was purified. The analysis of these complexes suggests the sequence of the assembly/disassembly process. It is shown that PSI-LHCI of C. reinhardtii is larger but far less stable than the complex from higher plants. Lhca2 and Lhca9 (the red-most antenna complexes), although present in the largest complex in 1:1 ratio with the core, are only loosely associated with it. This can explain the large variation in antenna composition of PSI-LHCI from C. reinhardtii found in the literature. The analysis of several subcomplexes with reduced antenna size allows determination of the position of Lhca2 and Lhca9 and leads to a proposal for a model of the organization of the Lhcas within the PSI-LHCI supercomplex.
在这项工作中,我们将莱茵衣藻的光系统 I(PSI)复合物纯化至均质。生化、蛋白质组学、光谱学和结构分析揭示了这个 PSI-LHCI 超复合物的主要性质。数据表明,最大的纯化复合物由一个核心复合物和九个 Lhca 天线组成,并且它包含所有 Lhca 基因产物。通过电子显微镜获得的 15Å分辨率的投影图显示,Lhcas 组织在核心的一侧呈双层半环排列,与之前的建议相反。一系列稳定的 PSI-LHCI 解体中间产物被纯化。对这些复合物的分析表明了组装/解体过程的顺序。结果表明,莱茵衣藻的 PSI-LHCI 虽然较大,但远不如高等植物的复合物稳定。Lhca2 和 Lhca9(最红的天线复合物)虽然与核心的比例为 1:1,但与核心的结合非常松散。这可以解释文献中发现的莱茵衣藻 PSI-LHCI 天线组成的巨大变化。对几个天线尺寸减小的亚复合物的分析允许确定 Lhca2 和 Lhca9 的位置,并提出了一个 PSI-LHCI 超复合物中 Lhcas 组织的模型。