Retinol-induced morphological changes of cultured bovine endothelial cells are accompanied by a marked increase in transglutaminase.
Retinol-induced morphological changes of cultured bovine endothelial cells are accompanied by a marked increase in transglutaminase.
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视黄醇诱导的培养牛内皮细胞的形态变化伴随着转谷氨酰胺酶的显着增加。
DOI：
10.1016/s0021-9258(19)47302-x
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
S. Hirose
中科院分区：
文献类型：
--
作者：
Kiyomitsu Nara;K. Nakanishi;H. Hagiwara;K. Wakita;S. Kojima;S. Hirose
视黄醇，形态发生素，已被证明诱导血管内皮细胞的形态变化，伴随着急性和特定的积累的80-kDa的蛋白质，这种视黄醇诱导的蛋白质（RIP）的纯化和表征已经揭示，它是一种转氨酶。培养牛颈动脉内皮细胞，用视黄醇处理，并检查形态学和蛋白质谱的变化。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析的提取物制备的视黄醇处理的细胞，经历了显着的变化的形状（从鹅卵石样的纺锤形）表明，视黄醇诱导的形态变化是伴随着显着增加的80 kDa的蛋白质。维甲酸也可引起类似的变化。通过阴离子交换和羟基磷灰石柱层析纯化80-kDa RIP。纯化的RIP的胰蛋白酶片段的氨基酸序列分析显示，牛RIP和豚鼠肝转氨酶的序列之间具有高度的同源性，这表明RIP是一种转氨酶。这是证实了活性测量; RIP表现出转氨酶活性，抗血清RIP免疫沉淀的活动。这些结果表明，转氨酶在维持形态和控制内皮细胞功能中起重要作用。
Retinol, a morphogen, has been shown to induce morphological changes in vascular endothelial cells, accompanied by an acute and specific accumulation of an 80-kDa protein; purification and characterization of this retinol-induced protein (RIP) have revealed that it is a transglutaminase. Endothelial cells from bovine carotid artery were cultured, treated with retinol, and examined for changes in morphology and protein profiles. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of extracts prepared from retinol-treated cells which had undergone a remarkable change in shape (from a cobblestone-like to a spindle-like shape) indicated that the retinol-induced morphological change is accompanied by a marked increase of an 80-kDa protein. Similar changes were also induced by retinoic acid. The 80-kDa RIP was purified by anion exchange and hydroxyapatite column chromatography. Amino acid sequencing of tryptic fragments of the purified RIP revealed a high degree of homology between the sequence of bovine RIP and that of guinea pig liver transglutaminase, suggesting that RIP is a transglutaminase. This was confirmed by activity measurements; RIP exhibited transglutaminase activity, and an antiserum against RIP immunoprecipitated the activity. These results suggest that transglutaminase plays important roles in the maintenance of morphology and the control of endothelial cell functions.