Swietnicki Wieslaw
Integrated Toxicology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702, USA.
Curr Opin Biotechnol. 2006 Aug;17(4):367-72. doi: 10.1016/j.copbio.2006.05.011. Epub 2006 Jun 5.
The successful expression and purification of proteins in an active form is essential for structural and biochemical studies. With rapid advances in genome sequencing and high-throughput structural biology, an increasing number of proteins are being identified as potential drug targets but are difficult to obtain in a form suitable for structural or biochemical studies. Although prokaryotic recombinant expression systems are often used, proteins obtained in this way are typically found to be insoluble. Several experimental approaches have therefore been developed to refold these aggregated proteins into a biologically active form, often suitable for structural studies. The major refolding strategies adopt one of two approaches - chromatographic methods or refolding in free solution - and both routes have been successfully used to refold a range of proteins. Future advances are likely to involve the development of automated approaches for protein refolding and purification.
以活性形式成功表达和纯化蛋白质对于结构和生化研究至关重要。随着基因组测序和高通量结构生物学的快速发展，越来越多的蛋白质被确定为潜在的药物靶点，但难以获得适合结构或生化研究的形式。尽管经常使用原核重组表达系统，但以这种方式获得的蛋白质通常是不溶性的。因此，已经开发了几种实验方法来将这些聚集的蛋白质重折叠成生物活性形式，通常适用于结构研究。主要的重折叠策略采用两种方法之一——色谱法或在自由溶液中重折叠——并且这两种途径都已成功用于重折叠一系列蛋白质。未来的进展可能涉及开发用于蛋白质重折叠和纯化的自动化方法。
