Berkmen Mehmet
New England Biolabs, 240 County Road, Ipswich, MA 01938, USA.
Protein Expr Purif. 2012 Mar;82(1):240-51. doi: 10.1016/j.pep.2011.10.009. Epub 2011 Nov 7.
Disulfide bonds are covalent bonds formed post-translationally by the oxidation of a pair of cysteines. A disulfide bond can serve structural, catalytic, and signaling roles. However, there is an inherent problem to the process of disulfide bond formation: mis-pairing of cysteines can cause misfolding, aggregation and ultimately result in low yields during protein production. Recent developments in the understanding of the mechanisms involved in the formation of disulfide bonds have allowed the research community to engineer and develop methods to produce multi-disulfide-bonded proteins to high yields. This review attempts to highlight the mechanisms responsible for disulfide bond formation in Escherichia coli, both in its native periplasmic compartment in wild-type strains and in the genetically modified cytoplasm of engineered strains. The purpose of this review is to familiarize the researcher with the biological principles involved in the formation of disulfide-bonded proteins with the hope of guiding the scientist in choosing the optimum expression system.
二硫键是一对半胱氨酸经氧化后在翻译后形成的共价键。二硫键可发挥结构、催化和信号传导作用。然而,二硫键形成过程存在一个固有问题:半胱氨酸错配会导致错误折叠、聚集,并最终在蛋白质生产过程中导致产量低下。对二硫键形成所涉及机制的最新认识进展,使研究界能够设计和开发出高产多二硫键结合蛋白的方法。本综述试图突出大肠杆菌中二硫键形成的机制,包括野生型菌株天然周质区室中的机制以及工程菌株经基因改造的细胞质中的机制。本综述的目的是让研究人员熟悉二硫键结合蛋白形成所涉及的生物学原理,以期指导科学家选择最佳表达系统。