CELLULAR INTERACTIONS AND TUBULIN DETYROSINATION IN FIBROBLASTIC AND EPITHELIAL-CELLS
CELLULAR INTERACTIONS AND TUBULIN DETYROSINATION IN FIBROBLASTIC AND EPITHELIAL-CELLS
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DOI：
10.1016/0248-4900(91)90061-q
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发表时间：
1991-01-01
影响因子：
2.7
通讯作者：
KARSENTI, E
中科院分区：
文献类型：
--
作者：
BRE, MH;PEPPERKOK, R;KARSENTI, E
在哺乳动物细胞中，大多数微管富含酪氨酸- α -微管蛋白（酪氨酸-微管蛋白）。微管的其他亚类以不同的数量存在，其中一些富含去酪氨酸α -微管蛋白（葡微管蛋白）。我们研究了细胞间相互作用对微管中微管蛋白水平的影响。利用抗酪氨酸微管蛋白和葡微管蛋白的抗体进行定量免疫荧光研究。我们发现，在已经建立细胞间接触的细胞中，glu-/ tyr1 -微管蛋白的比例高于分离细胞。我们还利用Schulze和Kirschner bbb的抗体阻断方法检测了细胞间相互作用对glu-/ tyr1 -微管蛋白比率的影响。微管主要含有tyr1 -微管蛋白，首先被一种针对tyr1 -微管蛋白的多克隆抗体和几层二抗阻断。然后用抗α -微管蛋白的单克隆抗体标记未阻断的微管。由于抗酪氨酸微管蛋白对微管的包覆效率取决于每个微管中酪氨酸微管蛋白的含量，因此该方法可以可视化每个细胞特定区域中酪氨酸微管蛋白富集或缺失的微管。微管在亚融合细胞中比在融合细胞中被更广泛地阻断，并且优先在细胞质的周围。在融合单层人工创面边缘的细胞中，阻塞微管的数量随时间缓慢增加（在2至4小时之间）。这些结果与细胞间接触导致成纤维细胞和上皮细胞微管蛋白去酪氨酸增加的假设是一致的。
In mammalian cells most microtubules are enriched in tyrosinated-alpha-tubulin (tyr-tubulin). Other subclasses of microtubules are present in variable amounts and some are enriched in detyrosinated-alpha-tubulin (glu-tubulin). We examined the effect of cell-cell interactions on the level of glu-tubulin in microtubules. This was studied by quantitative immunofluorescence using antibodies against tyr- and glu-tubulin. We found that in cells which have established cell-cell contacts, the ratio of glu-/tyr-tubulin is higher than in isolated cells. We also examined the effect of cell-cell interactions on the glu-/tyr-tubulin ratio by using the antibody blocking method of Schulze and Kirschner [42]. Microtubules containing mainly tyr-tubulin had been blocked first by a polyclonal antibody against tyr-tubulin and several layers of secondary antibodies. The unblocked microtubules were then labeled by a monoclonal antibody against alpha-tubulin. Since the coating efficiency of microtubules by the anti-tyr tubulin depends on the amount of tyr-tubulin in each microtubule, this procedure allows the visualization of microtubules enriched or depleted in tyr-tubulin in specific domains of each cell. Microtubules were more extensively blocked in subconfluent than in confluent cells and preferentially at the periphery of the cytoplasm. In cells present at the margin of an artificial wound produced in a confluent monolayer, the amount of blocked microtubules increased slowly with time (between 2 and 4 h). These results are consistent with the hypothesis that cell-cell contacts lead to increased tubulin detyrosination both in fibroblastic and epithelial cells.