改变大肠杆菌麦芽糖结合蛋白开放构象与闭合构象之间平衡的突变会阻碍其增强客蛋白溶解度的能力。
Mutations that alter the equilibrium between open and closed conformations of Escherichia coli maltose-binding protein impede its ability to enhance the solubility of passenger proteins.
作者信息
Nallamsetty Sreedevi, Waugh David S
机构信息
Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute at Frederick, P.O. Box B, Frederick, MD 21702-1201, USA.
出版信息
Biochem Biophys Res Commun. 2007 Dec 21;364(3):639-44. doi: 10.1016/j.bbrc.2007.10.060. Epub 2007 Oct 22.
Abstract
Certain highly soluble proteins, such as Escherichia coli maltose-binding protein (MBP), have the ability to enhance the solubility of their fusion partners, making them attractive vehicles for the production of recombinant proteins, yet the mechanism of solubility enhancement remains poorly understood. Here, we report that the solubility-enhancing properties of MBP are dramatically affected by amino acid substitutions that alter the equilibrium between its "open" and "closed" conformations. Our findings indicate that the solubility-enhancing activity of MBP is mediated by its open conformation and point to a likely role for the ligand-binding cleft in the mechanism of solubility enhancement.
摘要
某些高度可溶的蛋白质，如大肠杆菌麦芽糖结合蛋白（MBP），具有增强其融合伙伴溶解度的能力，使其成为生产重组蛋白的有吸引力的载体，然而溶解度增强的机制仍知之甚少。在这里，我们报告MBP的溶解度增强特性受到氨基酸取代的显著影响，这些取代改变了其“开放”和“封闭”构象之间的平衡。我们的研究结果表明，MBP的溶解度增强活性是由其开放构象介导的，并指出配体结合裂隙在溶解度增强机制中可能发挥的作用。
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