Antigen-induced secretion of histamine and the phosphorylation of myosin by protein kinase C in rat basophilic leukemia cells.
Antigen-induced secretion of histamine and the phosphorylation of myosin by protein kinase C in rat basophilic leukemia cells.
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大鼠嗜碱性白血病细胞中抗原诱导的组胺分泌和蛋白激酶 C 磷酸化肌球蛋白。
DOI：
10.1016/s0021-9258(18)63885-2
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
R. Adelstein
中科院分区：
文献类型：
--
作者：
R. Ludowyke;I. Peleg;M. Beaven;R. Adelstein
IgE介导的大鼠嗜碱性白血病（RBL-2 H3）细胞刺激导致组胺分泌。从这些细胞中免疫沉淀的肌球蛋白显示掺入其重链（200 kDa）和轻链（20 kDa）的放射性磷酸盐的量增加。在未受刺激的细胞中，肌球蛋白轻链的胰蛋白酶肽的二维映射揭示了一个磷酸肽，其含有被肌球蛋白轻链激酶磷酸化的丝氨酸残基。在刺激后，出现第二个磷酸肽，其含有被蛋白激酶C磷酸化的丝氨酸残基。来自肌球蛋白重链的胰蛋白酶磷酸肽图谱显示，未受刺激的细胞含有三种主要的磷酸肽。刺激后，一个新的胰蛋白酶磷酸肽出现含有丝氨酸位点磷酸化的蛋白激酶C.磷酸化的肌球蛋白轻链和重链的化学计量测定抗原刺激前后。在刺激前，肌球蛋白轻链含有0.4 mol磷酸盐/mol轻链，全部局限于未被蛋白激酶C磷酸化的丝氨酸。在10分钟内分泌其总组胺的44%的细胞表现出在由蛋白激酶C磷酸化的位点处的磷酸盐含量从0 mol磷酸盐/mol肌球蛋白亚基增加到0.7 mol磷酸盐/mol轻链和1 mol磷酸盐/mol重链。当用链球菌溶血素O使RBL-2 H3细胞具有可渗透性时，它们仍然显示出定性相似的分泌和磷酸化模式。我们的研究结果表明，组胺分泌的时间过程从刺激的RBL-2 H3细胞平行的肌球蛋白重链和轻链磷酸化的蛋白激酶C。
IgE-mediated stimulation of rat basophilic leukemia (RBL-2H3) cells results in the secretion of histamine. Myosin immunoprecipitated from these cells shows an increase in the amount of radioactive phosphate incorporated into its heavy (200 kDa) and light (20 kDa) chains. In unstimulated cells two-dimensional mapping of tryptic peptides of the myosin light chain reveals one phosphopeptide containing the serine residue phosphorylated by myosin light chain kinase. Following stimulation a second phosphopeptide appears containing a serine residue phosphorylated by protein kinase C. Tryptic phosphopeptide maps derived from myosin heavy chains show that unstimulated cells contain three major phosphopeptides. Following stimulation a new tryptic phosphopeptide appears containing a serine site phosphorylated by protein kinase C.The stoichiometry of phosphorylation of the myosin light and heavy chains was determined before and after antigenic stimulation. Before stimulation, myosin light chains contained 0.4 mol of phosphate/mol of light chain all confined to a serine not phosphorylated by protein kinase C. Cells that secreted 44% of their total histamine in 10 min exhibited an increase in phosphate content at sites phosphorylated by protein kinase C from 0 mol of phosphate/mol of myosin subunit to 0.7 mol of phosphate/mol of light chain and to 1 mol of phosphate/mol of heavy chain. When RBL-2H3 cells were made permeable with streptolysin O they still showed a qualitatively similar pattern of secretion and phosphorylation. Our results show that the time course of histamine secretion from stimulated RBL-2H3 cells parallels that of myosin heavy and light chain phosphorylation by protein kinase C.