使用蛋白质微阵列发现小鼠PDZ域的定量蛋白质相互作用网络
Michael A Stiffler1, Viara P Grantcharova, Mark Sevecka
1Department of Chemistry and Chemical Biology, Harvard University, 12 Oxford Street, Cambridge, Massachusetts 02138, USA.
Journal of the American Chemical Society
|April 28, 2006
概括
这项研究引入了一个以域为中心的方法来量化蛋白质-蛋白质相互作用,创建一个可靠的蛋白质相互作用网络. 这种方法提高了系统生物学研究的准确性.
科学领域:
- 系统生物学 系统生物学
- 分子生物学分子生物学
- 生物物理学的生物物理.
背景情况:
- 蛋白与蛋白相互作用 (PPI) 对生物过程至关重要.
- 目前用于绘制PPI的实验方法通常会产生具有高错误率 (错误阳性/负性) 的定性数据.
- 准确,定量PPI网络对于理解复杂的生物系统至关重要.
研究的目的:
- 开发一个以域为中心,高通量方法,用于定量测量PPI.
- 为PDZ域-相互作用创建一个平衡分离常数 (K(D)) 的黄金标准数据集.
- 使用定量生物物理数据建立可靠的蛋白质相互作用网络.
主要方法:
- 利用以域为中心的方法测量PDZ域和的平衡分离常数 (K(D)).
- 采用光偏振测定用于定量结合测量.
- 开发了蛋白质微阵列,用于PDZ域-相互作用的高通量选.
- 产生了520个PDZ-组合的黄金标准数据集.
主要成果:
- 实现了中等到高亲和力相互作用 (K(D) ≤10μM的高通量识别,其中14%的假阳性和14%的假阴性率.
- 创建了一个定量PPI网络,它汇总了以前已知的85%的相互作用.
- 发现了新的生物物理相互作用,特别是在共同表达的蛋白质之间.
- 证明了蛋白质序列和功能之间的强烈相关性,支持相互作用网络的逐渐演变.
结论:
- 基于域/的策略有效地将高吞吐量与高保真性结合起来,用于定量PPI网络构建.
- 定量网络提供了比定性方法更准确的生物相互作用表示.
- 这些发现支持一个由小的增量变化驱动的网络演变模型,保持功能连贯性.
相关概念视频
02:04Protein-protein Interfaces
Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a polypeptide...
02:26Protein Networks
An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
02:26Protein Networks
An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...