Mathesius Ulrike, Imin Nijat, Natera Siria H A, Rolfe Barry G
ARC Centre of Excellence for Integrative Legume Research, and Genomic Interactions Group, Research School of Biological Sciences, Australian National University, Canberra, Australia.
Methods Mol Biol. 2003;236:395-414. doi: 10.1385/1-59259-413-1:395.
To understand the function of all the genes in an organism, one needs to know not only which genes are expressed, when, and where, but also what the protein end products are and under which conditions they accumulate in certain tissues. Proteomics aims at describing the whole protein output of the genome and complements transcriptomic and metabolomic studies. Proteomics depends on extracting, separating, visualizing, identifying, and quantifying the proteins and their interactions present in an organism or tissue at any one time. All of these stages have limitations. Therefore, it is, at present, impossible to describe the whole proteome of any organism. Plants might synthesize many thousands of proteins at one time, and the whole potentially synthesized proteome certainly exceeds the number of estimated genes for that genome. This occurs because the gene products of one gene can differ due to alternative splicing and a variety of possible posttranslational modifications. It is, therefore, essential to optimize every step towards detecting the whole proteome while realizing the limitations. We concentrate here on the most commonly used steps in high-throughput plant proteomics with the techniques we have found most reproducible and with the highest resolution and quality.
为了解生物体中所有基因的功能,人们不仅需要知道哪些基因在何时何地表达,还需要知道蛋白质终产物是什么以及它们在何种条件下在特定组织中积累。蛋白质组学旨在描述基因组的全部蛋白质产出,并补充转录组学和代谢组学研究。蛋白质组学依赖于在任何时刻提取、分离、可视化、鉴定和定量生物体或组织中存在的蛋白质及其相互作用。所有这些阶段都有局限性。因此,目前还不可能描述任何生物体的完整蛋白质组。植物可能同时合成数千种蛋白质,整个潜在合成的蛋白质组肯定超过该基因组估计的基因数量。出现这种情况是因为一个基因的基因产物可能由于可变剪接和各种可能的翻译后修饰而有所不同。因此,在认识到局限性的同时,优化检测整个蛋白质组的每一步至关重要。我们在此集中讨论高通量植物蛋白质组学中最常用的步骤,这些步骤采用了我们发现最具可重复性、分辨率和质量最高的技术。