Büchel Claudia, Wilhelm Christian, Wagner Volker, Mittag Maria
Institute of Molecular Biosciences, Goethe University Frankfurt, 60438 Frankfurt, Germany.
Institute of Biology, Department of Plant Physiology, University of Leipzig, 04103 Leipzig, Germany.
J Plant Physiol. 2017 Oct;217:38-43. doi: 10.1016/j.jplph.2017.06.007. Epub 2017 Jul 4.
Comparative proteome analysis of subcellular compartments like thylakoid membranes and their associated supercomplexes can deliver important in-vivo information on the molecular basis of physiological functions which go far beyond to that what can be learnt from transcriptional-based gene expression studies. For instance, the finding that light intensity influences mainly the relative stoichiometry of subunits could be obtained only by high resolution proteome analysis. The high sensitivity of LC-ESI-MS/MS based proteome analysis allows the determination of proteins in very small subfractions along with their non-labeled semi quantitative analysis. This provides insights in the protein-protein interactions of supercomplexes that are the operative units in intact cells. Here, we have focused on functional proteome approaches for the identification of microalgal light-harvesting complex proteins in chloroplasts and the eyespot in general and in detail for those of diatoms that are exposed to varying light conditions.
对类囊体膜及其相关超复合体等亚细胞区室进行比较蛋白质组分析，可以提供有关生理功能分子基础的重要体内信息，这些信息远远超出了基于转录的基因表达研究所能获得的信息。例如，只有通过高分辨率蛋白质组分析才能获得光强度主要影响亚基相对化学计量的这一发现。基于液相色谱 - 电喷雾串联质谱（LC-ESI-MS/MS）的蛋白质组分析的高灵敏度使得能够在非常小的亚组分中测定蛋白质，并进行非标记的半定量分析。这为完整细胞中的操作单元——超复合体的蛋白质 - 蛋白质相互作用提供了见解。在这里，我们专注于功能蛋白质组方法，以鉴定叶绿体和眼点中的微藻捕光复合体蛋白质，特别是针对暴露于不同光照条件下的硅藻的此类蛋白质进行详细鉴定。
