Vasoactive intestinal peptide stimulates protein phosphorylation in a colonic epithelial cell line.
Vasoactive intestinal peptide stimulates protein phosphorylation in a colonic epithelial cell line.
复制标题
血管活性肠肽刺激结肠上皮细胞系中的蛋白质磷酸化。
DOI：
10.1152/ajpgi.1987.253.3.g420
复制
发表时间：
1987
期刊：
影响因子：
--
通讯作者：
J. Cohn
中科院分区：
文献类型：
--
作者：
J. Cohn
用T84结肠上皮细胞系检测神经体液刺激离子转运过程中蛋白质的磷酸化。将生长在胶原涂层滤膜上的T84细胞单层安装在Ussing小室中，以测量血管活性肠肽刺激的离子转运。刺激8~10min后，刺激活性分泌量最大。用双向凝胶电泳法检测伴随刺激分泌的蛋白磷酸化事件，以从以前用32PI标记的单层中分离磷蛋白。在暴露于血管活性肠肽的8分钟内，检测到几个磷酸化事件，包括~(32)P掺入表观分子量分别为17,000，18,000，23,000和37,000的四种可溶性蛋白质中的两到五倍。同样的磷酸化反应也发生在二丁酰腺苷3‘，5’-环一磷酸(CAMP)刺激的单层，这表明cAMP介导了这些细胞内事件。本研究表明，蛋白质磷酸化的变化伴随着血管活性肠肽的分泌作用，提示T84细胞为研究这种磷酸化事件调节肠细胞离子转运的可能性提供了一个有用的模型。
The T84 colonic epithelial cell line was used to examine protein phosphorylation during neurohumoral stimulation of ion transport. T84 cell monolayers grown on collagen-coated filters were mounted in Ussing chambers to measure ion transport stimulated by vasoactive intestinal peptide. Maximal stimulation of active secretion occurred after 8-10 min of stimulation. Protein phosphorylation events accompanying stimulated secretion were detected using two-dimensional gel electrophoresis to resolve phosphoproteins from monolayers previously labeled using 32Pi. Within 8 min of exposure to vasoactive intestinal peptide, several phosphorylation events were detected, including a two- to fivefold increase in 32P incorporation into four soluble proteins with apparent molecular weights of 17,000, 18,000, 23,000, and 37,000. The same phosphorylation response occurs in monolayers stimulated by dibutyryl adenosine 3',5'-cyclic monophosphate (cAMP), suggesting that cAMP mediates these intracellular events. This study indicates that changes in protein phosphorylation accompany the secretory action of vasoactive intestinal peptide and suggests that T84 cells offer a useful model for studying the possibility that such phosphorylation events regulate enterocyte ion transport.