Malys Naglis, Wishart Jill A, Oliver Stephen G, McCarthy John E G
Manchester Centre for Integrative Systems Biology, The University of Manchester, Manchester, United Kingdom.
Methods Enzymol. 2011;500:197-212. doi: 10.1016/B978-0-12-385118-5.00011-6.
Proteins together with metabolites, nucleic acids, lipids, and other intracellular molecules form biological systems that involve networks of functional and physical interactions. To understand these interactions and the many other characteristics of proteins in the context of biochemical networks and systems biology, research aimed at studying medium and large sets of proteins is required. This either involves an investigation focused on individual protein activities in the mixture (e.g., cell extracts) or a protein characterization in the isolated form. This chapter provides an overview on the currently available resources and strategies for isolation of proteins from Saccharomyces cerevisiae. The use of standardized gene expression systems is discussed, and protein production protocols applied to the data generation pipeline for systems biology are described in detail.
蛋白质与代谢物、核酸、脂质及其他细胞内分子共同构成了涉及功能和物理相互作用网络的生物系统。为了在生化网络和系统生物学的背景下理解这些相互作用以及蛋白质的许多其他特性,需要开展旨在研究中等规模和大规模蛋白质组的研究。这要么涉及聚焦于混合物(如细胞提取物)中单个蛋白质活性的研究,要么涉及对分离形式的蛋白质进行表征。本章概述了目前可用于从酿酒酵母中分离蛋白质的资源和策略。讨论了标准化基因表达系统的使用,并详细描述了应用于系统生物学数据生成流程的蛋白质生产方案。