昆虫粘蛋白型糖蛋白：黑腹果蝇细胞和组织中O-糖基化表位的免疫检测
Insect mucin-type glycoprotein: immunodetection of the O-glycosylated epitope in Drosophila melanogaster cells and tissues.
作者信息
Kramerov A A, Mikhaleva E A, Rozovsky Ya M, Pochechueva T V, Baikova N A, Arsenjeva E L, Gvozdev V A
机构信息
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, Russia.
出版信息
Insect Biochem Mol Biol. 1997 Jun;27(6):513-21. doi: 10.1016/s0965-1748(97)00026-x.
A mucin-type glycoprotein (GP) from cultured embryonic cells of Drosophila melanogaster was isolated and used to raise monoclonal antibodies (MAbs). Epitope(s) recognized by MAbs were sensitive to the treatment by O-glycanase, which specifically cleaves off O-linked mucin-type Gal(beta 1,3)GalNAc disaccharide, representing the major part of the carbohydrate moiety of Drosophila GP. Using high-affinity MAbs against carbohydrate epitopes of the Drosophila mucin GP we demonstrated its accumulation in culture medium, as well as in cultured cells, which proved to be regulated by 20-hydroxyecdysone. Mucin GPs carrying Gal(beta 1,3)GalNAc disaccharide recognized by the MAbs were immunochemically localized in several Drosophila tissues of ectodermal, mesodermal and germ line origin, including epidermal and follicle cells capable of their secretion.
从黑腹果蝇培养的胚胎细胞中分离出一种粘蛋白型糖蛋白（GP），并用于制备单克隆抗体（MAb）。单克隆抗体识别的表位对O-聚糖酶处理敏感，O-聚糖酶能特异性切割掉O-连接的粘蛋白型Gal（β1,3）GalNAc二糖，这是果蝇GP碳水化合物部分的主要成分。使用针对果蝇粘蛋白GP碳水化合物表位的高亲和力单克隆抗体，我们证明了它在培养基以及培养细胞中的积累，事实证明这受20-羟基蜕皮激素调控。携带单克隆抗体识别的Gal（β1,3）GalNAc二糖的粘蛋白型糖蛋白在起源于外胚层、中胚层和生殖系的几个果蝇组织中进行了免疫化学定位，包括能够分泌它们的表皮细胞和卵泡细胞。
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