Optimized native gel systems for separation of thylakoid protein complexes: novel super- and mega-complexes
Optimized native gel systems for separation of thylakoid protein complexes: novel super- and mega-complexes
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DOI：
10.1042/bj20102155
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发表时间：
2011-10-15
影响因子：
4.1
通讯作者：
Aro, Eva-Mari
中科院分区：
文献类型：
--
作者：
Jarvi, Sari;Suorsa, Marjaana;Aro, Eva-Mari
基于凝胶的类囊体膜蛋白复合物的分析是一个有价值的工具，以监测光合机制的动态。Native-PAGE保留了蛋白质复合物的组分，通常也保留了蛋白质复合物的构象，从而能够分析其亚基组成。然而，由于各种各样的非变性PAGE和类囊体增溶系统，该领域的文献和实际实验有时会引起混淆。本文介绍了高等植物类囊体膜蛋白复合物的非变性聚丙烯酰胺凝胶电泳分离的优化方法，主要包括：（i）分离植物类囊体膜蛋白复合物;去污剂的使用;（ii）增溶缓冲液的使用;和（iii）凝胶电泳法。特别注意的是从拟南芥叶片中分离高分子量的类囊体膜超级和巨型复合物。报道了几种新型的超级和巨型复合物，包括PS（光系统）I，PSII和LHC（捕光复合物）的各种组合。
Gel-based analysis of thylakoid membrane protein complexes represents a valuable tool to monitor the dynamics of the photosynthetic machinery. Native-PAGE preserves the components and often also the conformation of the protein complexes, thus enabling the analysis of their subunit composition. Nevertheless, the literature and practical experimentation in the field sometimes raise confusion owing to a great variety of native-PAGE and thylakoid-solubilization systems. In the present paper, we describe optimized methods for separation of higher plant thylakoid membrane protein complexes by native-PAGE addressing particularly: (i.) the use of detergent; (ii) the use of solubilization buffer; and (iii) the gel electrophoresis method. Special attention is paid to separation of high-molecular-mass thylakoid membrane super- and mega-complexes from Arabidopsis thaliana leaves. Several novel super- and mega-complexes including PS (photosystem) I, PSII and LHCs (light-harvesting complexes) in various combinations are reported.