Auerbach Daniel, Thaminy Safia, Hottiger Michael O, Stagljar Igor
Dualsystems Biotech, Zurich, Switzerland.
Proteomics. 2002 Jun;2(6):611-23. doi: 10.1002/1615-9861(200206)2:6<611::AID-PROT611>3.0.CO;2-Y.
The availability of completed genome sequences of several eukaryotic and prokaryotic species has shifted the focus towards the identification and characterization of all gene products that are expressed in a given organism. In order to cope with the huge amounts of data that have been provided by large-scale sequencing projects, high-throughout methodologies also need to be applied in the emerging field of proteomics. In this review, we discuss methods that have been recently developed in order to characterize protein interactions and their functional relevance on a large scale. We then focus on those methodologies that are suitable for the identification and characterization of protein-protein interactions, namely the yeast two-hybrid system and related methods. Several recent studies have demonstrated the power of automated approaches involving the yeast two-hybrid system in building so-called "interaction networks", which hold the promise of identifying the entirety of all interactions that take place between proteins expressed in a certain cell type or organism. We compare the yeast two-hybrid system with several other screening methods that have been developed to investigate interactions between proteins that are not amenable to conventional yeast two-hybrid screenings, such as transcriptional activators and integral membrane proteins. The eventual adaptation of such methods to a high-throughput format and their use in combination with automated yeast two-hybrid screenings will help in elucidating protein-protein interactions on a scale that would have been unthinkable just a few years ago.
几种真核生物和原核生物完整基因组序列的可得性，已将研究重点转向对特定生物体中表达的所有基因产物的识别和表征。为了处理大规模测序项目所提供的海量数据，高通量方法也需要应用于新兴的蛋白质组学领域。在这篇综述中，我们讨论了最近为大规模表征蛋白质相互作用及其功能相关性而开发的方法。然后，我们重点关注那些适用于识别和表征蛋白质-蛋白质相互作用的方法，即酵母双杂交系统及相关方法。最近的几项研究证明了涉及酵母双杂交系统的自动化方法在构建所谓“相互作用网络”方面的强大作用，这有望识别在特定细胞类型或生物体中表达的蛋白质之间发生的所有相互作用的整体情况。我们将酵母双杂交系统与其他几种已开发的筛选方法进行比较，这些方法用于研究不适用于传统酵母双杂交筛选的蛋白质之间的相互作用，如转录激活因子和整合膜蛋白。最终将这些方法调整为高通量形式，并将其与自动化酵母双杂交筛选结合使用，将有助于在几年前还无法想象的规模上阐明蛋白质-蛋白质相互作用。
