Glycoproteins and glycosaminoglycans synthesized by human keratinocytes in culture. Their role in cell-substratum adhesion.
Glycoproteins and glycosaminoglycans synthesized by human keratinocytes in culture. Their role in cell-substratum adhesion.
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由人角质形成细胞在培养物中合成的糖蛋白和糖胺聚糖。
DOI：
10.1042/bj2120355
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
L. Jenner
中科院分区：
文献类型：
--
作者：
G. Roberts;L. Jenner
提取人角质形成细胞在含有D-[1- 14 C]氨基葡萄糖的培养基中合成的糖蛋白和蛋白聚糖，并在十二烷基硫酸钠存在下通过聚丙烯酰胺凝胶电泳进行分析。用0.5%Triton X-100提取标记的角质形成细胞，除去大部分的糖缀合物，留下粘附于基质的细胞骨架和核残基。除了细胞骨架蛋白，有一个相对简单的配置文件的糖蛋白和糖胺聚糖与这种粘附的细胞骨架。这些糖蛋白由8种糖蛋白组成，分子量为10000。范围99000-232000，角蛋白区域中的5种蛋白质（mol. 42000-61000）、透明质酸和硫酸化糖胺聚糖。用半乳糖氧化酶（有或没有神经氨酸酶）/KB 3 H4的角质形成细胞的表面标记揭示了许多糖蛋白暴露在细胞表面上。通过研究在含有衣霉素的培养基中孵育后的糖蛋白和蛋白聚糖的表达以及用胰蛋白酶和透明质酸酶消化后的降解来检查糖蛋白和蛋白聚糖在将角质形成细胞附着于基质中的重要性。这些研究，连同通过用0.5%Triton X-100随后用0.2%十二烷基硫酸钠连续提取释放的糖缀合物的检查，揭示了摩尔重量的糖蛋白。232000在介导角质形成细胞附着于基质中具有重要作用。
Glycoproteins and proteoglycans synthesized by human keratinocytes in medium containing D-[1-14C]glucosamine were extracted and analysed by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. Extraction of the labelled keratinocytes with 0.5% Triton X-100 removed most of the glycoconjugates and left the cytoskeleton and nuclear residue adherent to the substratum. In addition to the cytoskeletal proteins, there was a relatively simple profile of glycoproteins and glycosaminoglycans associated with this adherent cytoskeleton. These consisted of eight glycoproteins in the mol.wt. range 99000-232000, five proteins in the keratin region (mol.wt. 42000-61000), hyaluronic acid and a sulphated glycosaminoglycan. Surface labelling of the keratinocytes with galactose oxidase (with or without neuraminidase)/KB3H4 revealed that many of the glycoproteins were exposed on the cell surface. The importance of the glycoproteins and proteoglycans in attaching the keratinocytes to the substratum was examined by studying their expression after incubation in medium containing tunicamycin and their degradation after digestion with trypsin and hyaluronidase. These studies, together with an examination of the glycoconjugates released by sequential extraction with 0.5% Triton X-100 followed by 0.2% sodium dodecyl sulphate, revealed that the glycoprotein of mol.wt. 232000 has an important role in mediating the attachment of keratinocytes to the substratum.