Large-scale screening for novel low-affinity extracellular protein interactions
Large-scale screening for novel low-affinity extracellular protein interactions
复制标题
DOI：
10.1101/gr.7187808
复制
发表时间：
2008-04-01
期刊：
影响因子：
7
通讯作者：
Wright, Gavin J.
中科院分区：
文献类型：
--
作者：
Bushell, K. Mark;Sollner, Christian;Wright, Gavin J.
细胞外蛋白质-蛋白质相互作用对于多细胞生物中的细胞间通讯和凝聚都是必不可少的。大约五分之一的人类基因编码膜系或分泌蛋白质，但它们在最近的大规模蛋白质相互作用数据集中基本上不存在，使得当前的相互作用网络存在偏见和不完整。这种差异是由于流行的高通量的方法来检测细胞外的相互作用，因为膜蛋白和它们的相互作用的生物化学棘手的不适合。例如，细胞表面蛋白质含有不溶性疏水跨膜区，并且它们的细胞外相互作用通常是高度瞬时的，具有小于一秒的半衰期。为了大规模检测瞬时细胞外相互作用，我们开发了AVEXIS（基于亲合力的细胞外相互作用筛选），这是一种高通量测定法，克服了这些技术问题，可以检测非常短暂的相互作用（半衰期
Extracellular protein-protein interactions are essential for both intercellular communication and cohesion within multicellular organisms. Approximately a fifth of human genes encode membrane-tethered or secreted proteins, but they are largely absent from recent large-scale protein interaction datasets, making current interaction networks biased and incomplete. This discrepancy is due to the unsuitability of popular high-throughput methods to detect extracellular interactions because of the biochemical intractability of membrane proteins and their interactions. For example, cell surface proteins contain insoluble hydrophobic transmembrane regions, and their extracellular interactions are often highly transient, having half-lives of less than a second. To detect transient extracellular interactions on a large scale, we developed AVEXIS (avidity-based extracellular interaction screen), a high-throughput assay that overcomes these technical issues and can detect very transient interactions (half-lives