Changes in cell-surface fucose-containing glycopeptides and adhesion of cultured intestinal epithelial cells as a function of cell density.
Changes in cell-surface fucose-containing glycopeptides and adhesion of cultured intestinal epithelial cells as a function of cell density.
复制标题
细胞表面含岩藻糖糖肽的变化和培养的肠上皮细胞的粘附随细胞密度的变化。
DOI：
10.1042/bj2110075
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Herscovics,A
中科院分区：
文献类型：
--
作者：
Sasak,W;Quaroni,A;Herscovics,A
融合培养的肠上皮细胞显示出更大的粘附比subconfluent细胞的基质。用[3 H]岩藻糖标记亚融合和融合细胞24小时，并通过温和的链霉蛋白酶处理释放细胞表面组分。在广泛的链霉蛋白酶消化后，在Bio-Gel P-6上分级分离细胞表面和细胞残留的糖肽。细胞表面含有比残留物更高比例的低分子量糖肽。未发现亚融合和融合细胞的总细胞表面糖肽的洗脱模式有显著差异。然而，融合细胞含有几乎两倍的[3 H]-岩藻糖标记的糖肽，其与伴刀豆球蛋白A-Sepharose结合，随后用20 mM-甲基α-D-吡喃葡萄糖苷洗脱，作为亚融合细胞。当结合的糖肽在Bio-Gel P-6上进行色谱分析时，发现融合细胞比亚融合细胞含有更大比例的低分子量糖肽。用唾液酸酶处理糖肽后，这种大小差异消除了。当亚融合细胞的生长被抑制与无毒浓度的视黄酸，[3 H]岩藻糖标记的糖肽的洗脱模式没有显着的影响，观察到无论是生物凝胶P-6或刀豆球蛋白A-琼脂糖。通过二维凝胶电泳，未发现亚融合细胞和融合细胞的总[3 H]岩藻糖标记糖蛋白有显著差异。这表明，[3 H]岩藻糖标记的糖肽亚融合和融合细胞之间的差异是细胞密度依赖性，而不是生长依赖性，这些差异可能是由于糖基化机制的一些变化。此外，细胞表面糖肽的差异可能与细胞对基质粘附的变化有关。
Confluent cultured intestinal epithelial cells displayed greater adhesion to the substratum than did subconfluent cells. Subconfluent and confluent cells were labelled with [3H]fucose for 24h and the cell-surface components were released by mild Pronase treatment. After extensive Pronase digestion, cell-surface and cell-residue glycopeptides were fractionated on Bio-Gel P-6. The cell surface contained a higher proportion of lower-molecular-weight glycopeptides than the residue. No significant difference in elution pattern was found between total cell-surface glycopeptides of subconfluent and confluent cells. However, confluent cells contained almost twice as much [3H]-fucose-labelled glycopeptides that were bound to concanavalin A-Sepharose and were subsequently eluted with 20mM-methyl alpha-D-glucopyranoside as subconfluent cells. When the bound glycopeptides were chromatographed on Bio-Gel P-6, it was found that confluent cells contained a larger proportion of lower-molecular-weight glycopeptides than subconfluent cells. This difference in size was eliminated after treatment of glycopeptides with sialidase. When growth of subconfluent cells was inhibited with a non-toxic concentration of retinoic acid, no significant effect on the elution pattern of [3H]fucose-labelled glycopeptides was observed on either Bio-Gel P-6 or concanavalin A-Sepharose. No significant difference was found in the total [3H]fucose-labelled glycoproteins from subconfluent and confluent cells by two-dimensional gel electrophoresis. It is suggested that the differences in [3H]fucose-labelled glycopeptides between subconfluent and confluent cells are cell-density-dependent rather than growth-dependent, and that these differences are likely to result from some changes in glycosylation mechanism(s). Furthermore, the differences in cell-surface glycopeptides may be related to the changes in the adhesion of the cells to the substratum.