Division of Cell Recognition Study, Institute of Molecular Biomembrane and Glycobiology, Tohoku Medical and Pharmaceutical University, Sendai, Japan.
Methods Mol Biol. 2020;2132:359-367. doi: 10.1007/978-1-0716-0430-4_35.
SUEL-like lectins, also termed rhamnose-binding lectins (RBL), are unique in animal lectin families because of their tandemly repeated structure that is characteristic of carbohydrate-recognition domains, as well as their α-galactoside-binding capacity. RBLs are known to be expressed in inclusion bodies in Escherichia coli. Here, we describe the methods for the expression and refolding of Silurus asotus lectin (SAL) using E. coli KRX as the host strain. From our results, highly basic and reduced conditions due to arginine and dithiothreitol, respectively, tend to keep SAL recombinants soluble.
SUEL 样凝集素,也称为鼠李糖结合凝集素 (RBL),在动物凝集素家族中是独特的,因为它们串联重复的结构是碳水化合物识别结构域的特征,以及它们的 α-半乳糖苷结合能力。RBL 已知在大肠杆菌的包涵体中表达。在这里,我们描述了使用大肠杆菌 KRX 作为宿主菌株表达和重折叠鲇鱼凝集素 (SAL)的方法。根据我们的结果,由于精氨酸和二硫苏糖醇分别导致的高度碱性和还原条件有利于保持 SAL 重组蛋白的可溶性。