Stein Viktor
Fachbereich Biologie, Technische Universität Darmstadt, 64287, Darmstadt, Germany.
Methods Mol Biol. 2017;1596:3-25. doi: 10.1007/978-1-4939-6940-1_1.
Synthetic protein switches with tailored response functions are finding increasing applications as tools in basic research and biotechnology. With a number of successful design strategies emerging, the construction of synthetic protein switches still frequently necessitates an integrated approach that combines detailed biochemical and biophysical characterization in combination with high-throughput screening to construct tailored synthetic protein switches. This is increasingly complemented by computational strategies that aim to reduce the need for costly empirical optimization and thus facilitate the protein design process. Successful computational design approaches range from analyzing phylogenetic data to infer useful structural, biophysical, and biochemical information to modeling the structure and function of proteins ab initio. The following chapter provides an overview over the theoretical considerations and experimental approaches that have been successful applied in the construction of synthetic protein switches.
具有定制响应功能的合成蛋白开关作为基础研究和生物技术中的工具,其应用越来越广泛。随着多种成功设计策略的出现,合成蛋白开关的构建仍常常需要一种综合方法,即将详细的生化和生物物理表征与高通量筛选相结合,以构建定制的合成蛋白开关。旨在减少昂贵的经验优化需求从而促进蛋白质设计过程的计算策略越来越多地对其起到补充作用。成功的计算设计方法涵盖从分析系统发育数据以推断有用的结构、生物物理和生化信息到从头开始模拟蛋白质的结构和功能。以下章节概述了在合成蛋白开关构建中成功应用的理论考量和实验方法。