Shida H, Gorbsky G, Shida M, Steinberg M S
J Cell Biochem. 1982;20(2):113-26. doi: 10.1002/jcb.240200204.
Correlated ultrastructural and biochemical methods were used to identify and localize Concanavalin A (Con A) receptors in the desmosomes of bovine epidermis. Specific carbohydrate residues were labeled with ferritin-Con A in thin sections of tissue embedded in a hydrophilic resin. Quantitative mapping of ferritin distribution in labeled desmosomes revealed that Con A receptors are localized in the intercellular zone and concentrated along the desmosomal midline or central dense stratum. Labeling was almost entirely absent when sections were treated with ferritin-Con A in the presence of 0.1 M alpha-methyl mannoside, a hapten-inhibitor of Con A. "Whole" desmosomes and desmosomal intercellular regions (desmosomal "cores") were purified from bovine muzzle epidermis. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis reveals a limited number of major desmosomal protein constituents. Certain of these are glycoproteins and are greatly enriched in the core fraction. Almost all the desmosomal glycoproteins are intensely labeled when electrophoretic gels of whole desmosome or core fractions are exposed to fluorescent Concanavalin A.
运用相关的超微结构和生化方法来鉴定和定位牛表皮桥粒中的伴刀豆球蛋白A(Con A)受体。在嵌入亲水性树脂的组织薄片中,用铁蛋白-Con A标记特定的碳水化合物残基。对标记桥粒中铁蛋白分布的定量绘图显示,Con A受体位于细胞间区域,并沿桥粒中线或中央致密层集中。当切片在0.1Mα-甲基甘露糖苷(Con A的半抗原抑制剂)存在的情况下用铁蛋白-Con A处理时,几乎完全没有标记。从牛口鼻表皮中纯化出“完整”的桥粒和桥粒细胞间区域(桥粒“核心”)。十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳显示桥粒主要蛋白质成分数量有限。其中某些是糖蛋白,并且在核心部分中大量富集。当完整桥粒或核心部分的电泳凝胶暴露于荧光伴刀豆球蛋白A时,几乎所有桥粒糖蛋白都被强烈标记。