Birungi Mariam, Folea Mihaela, Battchikova Natalia, Xu Min, Mi Hualing, Ogawa Teruo, Aro Eva-Mari, Boekema Egbert J
Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
Biochim Biophys Acta. 2010 Sep;1797(9):1681-6. doi: 10.1016/j.bbabio.2010.06.004. Epub 2010 Jun 12.
Cyanobacterial NDH-1 is a multisubunit complex involved in proton translocation, cyclic electron flow around photosystem I and CO2 uptake. The function and location of several of its small subunits are unknown. In this work, the location of the small subunits NdhL, -M, -N, -O and CupS of Synechocystis 6803 NDH-1 was established by electron microscopy (EM) and single particle analysis. To perform this, the subunits were enlarged by fusion with the YFP protein. After classification of projections, the position of the YFP tag was revealed; all five subunits are integrated in the membrane domain. The results on NDH-1 demonstrate that a GFP tag can be revealed after data processing of EM data sets of moderate size, thus showing that this way of labeling is a fast and reliable way for subunit mapping in multisubunit complexes after partial purification.
蓝藻NDH-1是一种多亚基复合体，参与质子转运、围绕光系统I的循环电子流以及二氧化碳的摄取。其几个小亚基的功能和定位尚不清楚。在这项工作中，通过电子显微镜（EM）和单颗粒分析确定了集胞藻6803 NDH-1的小亚基NdhL、-M、-N、-O和CupS的定位。为此，通过与YFP蛋白融合来扩大这些亚基。在对投影进行分类后，揭示了YFP标签位置；所有五个亚基都整合在膜结构域中。关于NDH-1的结果表明，在对中等规模的EM数据集进行数据处理后，可以揭示GFP标签，从而表明这种标记方式是部分纯化后多亚基复合体中亚基定位的一种快速且可靠的方法。
