Graduate School of Engineering, Nagasaki University, Nagasaki, Japan.
Methods Mol Biol. 2020;2132:159-164. doi: 10.1007/978-1-0716-0430-4_16.
CEL-III is a Ca-dependent and galactose-specific lectin purified from the sea cucumber, Cucumaria echinata; it exhibits hemolytic and hemagglutinating activities. CEL-III consists of the following three distinct domains: two N-terminal carbohydrate-binding domains (1 and 2), which adopt β-trefoil folds such as the B-chain of ricin and are members of the (QXW) motif family, and domain 3, an oligomerization domain. After binding to the cell surface carbohydrate chains through domains 1 and 2, domain 3 self-associates to form transmembrane pores composed of CEL-III heptamers, leading to cell lysis or death. In this chapter, the purification and carbohydrate-coated microplate binding assay of CEL-III are described.
CEL-III 是从海参中纯化得到的一种 Ca2+依赖性和半乳糖特异性凝集素,具有溶血和凝集活性。CEL-III 由以下三个不同的结构域组成:两个 N 端糖结合结构域(1 和 2),它们采用 β-三叶折叠,如蓖麻毒素的 B 链,属于(QXW)基序家族,和结构域 3,寡聚化结构域。通过结构域 1 和 2 与细胞表面的碳水化合物链结合后,结构域 3 自身缔合形成由 CEL-III 七聚体组成的跨膜孔,导致细胞裂解或死亡。在本章中,描述了 CEL-III 的纯化和糖包被微孔板结合测定。