Pasteurella multocida toxin, a potent mitogen, increases inositol 1,4,5-trisphosphate and mobilizes Ca2+ in Swiss 3T3 cells.
Pasteurella multocida toxin, a potent mitogen, increases inositol 1,4,5-trisphosphate and mobilizes Ca2+ in Swiss 3T3 cells.
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多杀性巴氏杆菌毒素是一种有效的有丝分裂原，可增加 Swiss 3T3 细胞中肌醇 1,4,5-三磷酸的含量并动员 Ca2+。
DOI：
10.1016/s0021-9258(19)67725-2
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发表时间：
1991
期刊：
影响因子：
--
通讯作者：
Enrique Rozengurt
中科院分区：
文献类型：
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作者：
J. Staddon;C. J. Barker;Anne C. Murphy;Neil Chanter;A. Lax;Robert H. Michell;Enrique Rozengurt
多杀性巴氏杆菌毒素，无论是原生的还是重组的，对于Swiss 3T3细胞来说都是一种非常有效的丝裂原，并能增强总肌醇磷酸的形成(Rozengurt， E., Higgins， T., Changer， N., Lax, a.j., and Staddon， J.M. (1990) Proc. Natl。学会科学。美国87,123-127)。多杀毒杆菌毒素也能刺激二酰基甘油的产生并激活蛋白激酶C (Staddon, j.m., Chanter， N., Lax, a.j., Higgins, t.e., and Rozengurt， E.（1990）。化学，265,11841-11848)。本文采用[3H]肌醇标记法和高效液相色谱法分析了重组多杀p.a毒素处理细胞中的肌醇磷酸。重组多杀p.a毒素刺激了[3H]肌醇1,4,5-三磷酸（[3H]Ins(1,4,5)P3）及其代谢产物，包括Ins(1,3,4,5)P4、Ins(1,3,4)P3、Ins(1,4)P2、Ins(4/5)P和Ins(1/3)P的增加。这些不同的肌醇磷酸盐的细胞含量的增加与bombesin引起的非常相似。此外，重组多杀假单胞菌毒素，如bombesin，动员细胞内的Ca2+池。重组P. multicida毒素预处理大大降低了bombesin的Ca2（+）动员作用，这与两种药物从共同池中动员Ca2+一致。重组多杀p.a multocida毒素对肌醇磷酸盐的增强和Ca2+的动员被溶酶体营养剂甲胺、氯化铵和氯喹阻断，并在剂量依赖滞后期发生。在去除细胞外毒素后，重组多杀假单胞菌毒素对肌醇磷酸产生的刺激持续存在，这与bombesin作用的可逆性相反。重组多杀假单胞菌毒素，不像炸弹蛋白和鸟苷5'- o -（γ -硫代三磷酸），不引起肌醇磷酸盐在渗透细胞中的释放。这些数据表明，重组多杀假单胞菌毒素在细胞内作用，刺激磷脂酶c介导的磷脂酰肌醇4,5-二磷酸水解。
Pasteurella multocida toxin, both native and recombinant, is an extremely potent mitogen for Swiss 3T3 cells and acts to enhance the formation of total inositol phosphates (Rozengurt, E., Higgins, T., Changer, N., Lax, A.J., and Staddon, J.M. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 123-127). P. multocida toxin also stimulates diacylglycerol production and activates protein kinase C (Staddon, J.M., Chanter, N., Lax, A.J., Higgins, T.E., and Rozengurt, E. (1990) J. Biol. Chem. 265, 11841-11848). Here we analyze, by [3H]inositol labeling and high performance liquid chromatography, the inositol phosphates in recombinant P. multocida toxin-treated cells. Recombinant P. multocida toxin stimulated increases in [3H]inositol 1,4,5-trisphosphate ([3H]Ins(1,4,5)P3) and its metabolic products, including Ins(1,3,4,5)P4, Ins(1,3,4)P3, Ins(1,4)P2, Ins(4/5)P, and Ins(1/3)P. The profile of the increase in the cellular content of these distinct inositol phosphates was very similar to that elicited by bombesin. Furthermore, recombinant P. multocida toxin, like bombesin, mobilizes an intracellular pool of Ca2+. Recombinant P. multocida toxin pretreatment greatly reduces the Ca2(+)-mobilizing action of bombesin, consistent with Ca2+ mobilization from a common pool by the two agents. The enhancement of inositol phosphates and mobilization of Ca2+ by recombinant P. multocida toxin were blocked by the lysosomotrophic agents methylamine, ammonium chloride, and chloroquine and occurred after a dose-dependent lag period. The stimulation of inositol phosphate production by recombinant P. multocida toxin persisted after removal of extracellular toxin, in contrast to the reversibility of the action of bombesin. Recombinant P. multocida toxin, unlike bombesin and guanosine 5'-O-(gamma-thiotriphosphate), did not cause the release of inositol phosphates in permeabilized cells. These data demonstrate that recombinant P. multocida toxin, acting intracellularly, stimulates the phospholipase C-mediated hydrolysis of phosphatidylinositol 4,5-bisphosphate.