EDEMs 的体外甘露糖苷酶活性测定：增强内质网降解的α-甘露糖苷酶样蛋白。
In Vitro Mannosidase Assay of EDEMs: ER Degradation-Enhancing α-Mannosidase-Like Proteins.
机构信息
Laboratory of Molecular and Cellular Biology, Institute for Frontier Life and Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
出版信息
Methods Mol Biol. 2020;2132:151-158. doi: 10.1007/978-1-0716-0430-4_15.
Quality control of newly synthesized glycoproteins is tightly regulated by sugar processing of N-glycans and by recognition of specific glycan structures by lectins in the endoplasmic reticulum (ER). Mannose trimming and its recognition determine the targeting of misfolded glycoproteins for ER-associated degradation. ER degradation-enhancing α-mannosidase-like (EDEM) proteins in mammals and their homologue Htm1p/Mnl1p in Saccharomyces cerevisiae are involved in this process. To analyze the function of EDEM proteins, we expressed and purified recombinant EDEM3 from HEK293 cells and assessed its mannose-trimming activity in vitro.
新合成的糖蛋白的质量控制受到内质网 (ER) 中 N-聚糖的糖加工和凝集素对特定糖结构的识别的严格调控。甘露糖修剪及其识别决定了错误折叠糖蛋白的靶向 ER 相关降解。哺乳动物中的 ER 降解增强的 α-甘露糖苷酶样 (EDEM) 蛋白及其在酿酒酵母中的同源物 Htm1p/Mnl1p 参与了这一过程。为了分析 EDEM 蛋白的功能，我们从 HEK293 细胞中表达和纯化了重组 EDEM3，并在体外评估了其甘露糖修剪活性。
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