Ford Kristina L, Chin Tony, Srivastava Vaibhav, Zeng Wei, Doblin Monika S, Bulone Vincent, Bacic Antony
Australian Research Council Centre of Excellence in Plant Cell Walls, School of BioSciences, The University of Melbourne, Victoria 3010, Australia.
Division of Glycoscience, School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Centre, 106 91 Stockholm, Sweden.
Proteomes. 2016 Jul 20;4(3):23. doi: 10.3390/proteomes4030023.
The Golgi apparatus (GA) is a crucial organelle in the biosynthesis of non-cellulosic polysaccharides, glycoproteins and proteoglycans that are primarily destined for secretion to the cell surface (plasma membrane, cell wall and apoplast). Only a small proportion of the proteins involved in these processes have been identified in plants, with the majority of their functions still unknown. The availability of a GA proteome would greatly assist plant biochemists, cell and molecular biologists in determining the precise function of the cell wall-related proteins. There has been some progress towards defining the GA proteome in the model plant system , yet in commercially important species, such as either the cereals or woody species there has been relatively less progress. In this study, we applied discontinuous sucrose gradient centrifugation to partially enrich GA from suspension cell cultures (SCCs) and combined this with stable isotope labelling (iTRAQ) to determine protein sub-cellular locations. Results from a representative grass species, Italian ryegrass () and a dicot species, black cottonwood () are compared. The results confirm that membrane fractionation approaches that provide effective GA-enriched fractions for proteomic analyses in are much less effective in the species examined here and highlight the complexity of the GA, both within and between species.
高尔基体（GA）是生物合成非纤维素多糖、糖蛋白和蛋白聚糖的关键细胞器，这些物质主要用于分泌到细胞表面（质膜、细胞壁和质外体）。在植物中，参与这些过程的蛋白质只有一小部分已被鉴定出来，它们的大多数功能仍然未知。高尔基体蛋白质组的可得性将极大地帮助植物生物化学家、细胞和分子生物学家确定细胞壁相关蛋白质的精确功能。在模式植物系统中，在定义高尔基体蛋白质组方面已经取得了一些进展，然而在商业上重要的物种，如谷物或木本物种中，进展相对较少。在本研究中，我们应用不连续蔗糖梯度离心法从悬浮细胞培养物（SCCs）中部分富集高尔基体，并将其与稳定同位素标记（iTRAQ）相结合，以确定蛋白质的亚细胞定位。比较了一种代表性禾本科植物意大利黑麦草（Lolium multiflorum）和一种双子叶植物黑杨（Populus trichocarpa）的结果。结果证实，为拟南芥（Arabidopsis thaliana）蛋白质组分析提供有效高尔基体富集组分的膜分级分离方法在此处研究的物种中效果要差得多，并突出了物种内部和物种之间高尔基体的复杂性。
