Institute of Biomedical Chemistry (IBMC), 119121 Moscow, Russia.
Int J Mol Sci. 2022 Dec 10;23(24):15697. doi: 10.3390/ijms232415697.
Most proteins function as part of various complexes, forming via stable and dynamic protein-protein interactions (PPIs). The profiling of PPIs expands the fundamental knowledge about the structures, functions, and regulation patterns of protein complexes and intracellular molecular machineries. Protein interactomics aims at solving three main tasks: (1) identification of protein partners and parts of complex intracellular structures; (2) analysis of PPIs parameters (affinity, molecular-recognition specificity, kinetic rate constants, and thermodynamic-parameters determination); (3) the study of the functional role of novel PPIs. The purpose of this work is to update the current state and prospects of multi-omics approaches to profiling of proteins involved in the formation of stable complexes. Methodological paradigm includes a development of protein-extraction and -separation techniques from tissues or cellular lysates and subsequent identification of proteins using mass-spectrometry analysis. In addition, some aspects of authors' experimental platforms, based on high-performance size-exclusion chromatography, procedures of molecular fishing, and protein identification, as well as the possibilities of interactomic taxonomy of each protein, are discussed.
大多数蛋白质作为各种复合物的一部分发挥作用，通过稳定和动态的蛋白质-蛋白质相互作用（PPIs）形成。PPIs 的分析扩展了关于蛋白质复合物和细胞内分子机器的结构、功能和调节模式的基本知识。蛋白质相互作用组学旨在解决三个主要任务：（1）鉴定蛋白质伴侣和复杂细胞内结构的部分；（2）分析 PPIs 参数（亲和力、分子识别特异性、动力学速率常数和热力学参数确定）；（3）研究新的 PPIs 的功能作用。这项工作的目的是更新当前状态和多组学方法在研究稳定复合物形成中涉及的蛋白质的前景。方法学范例包括从组织或细胞裂解物中开发蛋白质提取和分离技术，然后使用质谱分析鉴定蛋白质。此外，还讨论了作者基于高性能凝胶过滤色谱、分子捕捞程序和蛋白质鉴定的实验平台的一些方面，以及每个蛋白质的相互作用组分类的可能性。
