Brysk M M, Rajaraman S, Penn P, Chen S J
Differentiation. 1986;32(3):230-7. doi: 10.1111/j.1432-0436.1986.tb00578.x.
We isolated a concanavalin A (Con-A)-binding glycoprotein from human stratum corneum by nonionic detergent extraction, lectin affinity chromatography, and preparative gel electrophoresis. This glycoprotein migrates as a single band at 40 kilodaltons at sodium-dodecyl-sulfate gel electrophoresis with or without the presence of 2-mercaptoethanol. It was shown to have a heterogeneous distribution between pH 5.6 and 7.6 by isoelectric focusing. The glycoprotein is histidine rich (10.4%) but is distinct from other histidine-rich proteins (epidermal filaggrin and the histidine-rich glycoprotein from serum). It does not bind to lectins specific for L-fucose or alpha-D-galactose. We prepared a monospecific polyclonal antibody to the 40-kilodalton glycoprotein; at the ultrastructural level, it cytoimmunolocalizes exclusively to the membranes of the stratum corneum. A unique feature of the glycoprotein is that it is an endogenous lectin: it hemagglutinates trypsinized and gluteraldehyde-fixed rabbit erythrocytes. The inhibition of its hemagglutination was found to be greatest with amino sugars, down to a saccharide concentration of 10(-5) mM. Such a high affinity of binding at the cell surface suggests that this glycoprotein is a major carbohydrate-binding, cross-linking molecule that holds adjacent corneocytes together in the stratum corneum. We hypothesize that this lectin plays a role in the adhesion and desquamation of the stratum corneum.
我们通过非离子去污剂提取、凝集素亲和层析和制备性凝胶电泳，从人角质层中分离出一种伴刀豆球蛋白A（Con-A）结合糖蛋白。在有或没有2-巯基乙醇存在的情况下，这种糖蛋白在十二烷基硫酸钠凝胶电泳中以40千道尔顿的单一条带迁移。通过等电聚焦显示，它在pH 5.6至7.6之间具有异质分布。该糖蛋白富含组氨酸（10.4%），但与其他富含组氨酸的蛋白质（表皮聚角蛋白和血清中的富含组氨酸糖蛋白）不同。它不与对L-岩藻糖或α-D-半乳糖特异的凝集素结合。我们制备了针对这种40千道尔顿糖蛋白的单特异性多克隆抗体；在超微结构水平上，它仅在角质层膜上进行细胞免疫定位。这种糖蛋白的一个独特特征是它是一种内源性凝集素：它能使经胰蛋白酶处理和戊二醛固定的兔红细胞发生血凝。发现其血凝抑制作用在氨基糖存在时最强，低至10^(-5) mM的糖浓度。在细胞表面如此高的结合亲和力表明，这种糖蛋白是一种主要的碳水化合物结合、交联分子，它将角质形成细胞在角质层中彼此连接在一起。我们推测这种凝集素在角质层的黏附与脱屑过程中起作用。
