Kobashigawa Yoshihiro, Kumeta Hiroyuki, Ogura Kenji, Inagaki Fuyuhiko
Department of Structural Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, Sapporo, Hokkaido, 060-0810, Japan.
J Biomol NMR. 2009 Mar;43(3):145-50. doi: 10.1007/s10858-008-9296-5. Epub 2009 Jan 13.
Sample solubility is essential for structural studies of proteins by solution NMR. Attachment of a solubility enhancement tag, such as GB1, MBP and thioredoxin, to a target protein has been used for this purpose. However, signal overlap of the tag with the target protein often made the spectral analysis difficult. Here we report a sortase-mediated protein ligation method to eliminate NMR signals arising from the tag by preparing the isotopically labeled target protein attached with the non-labeled GB1 tag at the C-terminus.
样品溶解度对于通过溶液核磁共振进行蛋白质结构研究至关重要。为此，人们已将诸如GB1、麦芽糖结合蛋白（MBP）和硫氧还蛋白等溶解度增强标签连接到目标蛋白上。然而，标签与目标蛋白的信号重叠常常使光谱分析变得困难。在此，我们报告一种分选酶介导的蛋白质连接方法，通过制备在C端连接有未标记GB1标签的同位素标记目标蛋白，来消除标签产生的核磁共振信号。
