Guruharsha K G, Obar Robert A, Mintseris Julian, Aishwarya K, Krishnan R T, Vijayraghavan K, Artavanis-Tsakonas Spyros
Department of Cell Biology; Harvard Medical School, Boston, MA, USA.
Fly (Austin). 2012 Oct-Dec;6(4):246-53. doi: 10.4161/fly.22108.
Proteins perform essential cellular functions as part of protein complexes, often in conjunction with RNA, DNA, metabolites and other small molecules. The genome encodes thousands of proteins but not all of them are expressed in every cell type; and expressed proteins are not active at all times. Such diversity of protein expression and function accounts for the level of biological intricacy seen in nature. Defining protein-protein interactions in protein complexes, and establishing the when, what and where of potential interactions, is therefore crucial to understanding the cellular function of any protein-especially those that have not been well studied by traditional molecular genetic approaches. We generated a large-scale resource of affinity-tagged expression-ready clones and used co-affinity purification combined with tandem mass-spectrometry to identify protein partners of nearly 5,000 Drosophila melanogaster proteins. The resulting protein complex "map" provided a blueprint of metazoan protein complex organization. Here we describe how the map has provided valuable insights into protein function in addition to generating hundreds of testable hypotheses. We also discuss recent technological advancements that will be critical in addressing the next generation of questions arising from the map.
蛋白质作为蛋白质复合物的一部分发挥着重要的细胞功能，通常与RNA、DNA、代谢物及其他小分子协同作用。基因组编码数千种蛋白质，但并非所有蛋白质都在每种细胞类型中表达；而且已表达的蛋白质也并非始终处于活性状态。蛋白质表达和功能的这种多样性解释了自然界中所见到的生物复杂性水平。因此，确定蛋白质复合物中的蛋白质-蛋白质相互作用，并确定潜在相互作用的时间、内容和位置，对于理解任何蛋白质的细胞功能至关重要，尤其是那些尚未通过传统分子遗传学方法得到充分研究的蛋白质。我们生成了大量带有亲和标签的可表达克隆资源，并使用共亲和纯化结合串联质谱法来鉴定近5000种黑腹果蝇蛋白质的蛋白质伴侣。由此产生的蛋白质复合物“图谱”提供了后生动物蛋白质复合物组织的蓝图。在此，我们描述了该图谱除了产生数百个可测试的假设之外，如何为蛋白质功能提供了有价值的见解。我们还讨论了近期的技术进步，这些进步对于解决由该图谱引发的下一代问题至关重要。
