Kudoh M, Nakamura T, Koyama J
Mol Immunol. 1985 Sep;22(9):1099-105. doi: 10.1016/0161-5890(85)90113-0.
As demonstrated by others, diisopropyl fluorophosphate (DFP) markedly inhibits the O2- generation from guinea-pig polymorphonuclear leukocytes (PMN) stimulated by an antibody complex with ovalbumin (Ag-Ab complex), and also the intracellular uptake of antibody-sensitized erythrocytes by the cells. However, when PMN were treated with DFP and washed to remove the inhibitor, they again became able to exhibit the O2- -generating and phagocytic activities. The [3H]DFP-labeling of intact PMN followed by solubilization with Triton N101, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed the existence of several [3H]DFP-labeled proteins with different mol. wts, which disappeared on pretreatment of cells with cold DFP. However, stimulation of DFP-pretreated PMN with Ag-Ab complex in the presence of [3H]DFP resulted in the appearance of a [3H]DFP-labeled, membrane-bound protein with a mol. wt of 40,000. This protein was isolated by affinity chromatography of the solubilized PMN and phagosomes on anti-Ig antibody-Sepharose 4B. Although the enzymatic properties of the protein are not clear, the results so far obtained suggest that it is a putative, stimulus-activated serine protease participating in the triggering events leading to the activation of NADPH oxidase responsible for the respiratory burst and the formation of phagosomes.
正如其他人所证明的，二异丙基氟磷酸酯（DFP）能显著抑制豚鼠多形核白细胞（PMN）由卵清蛋白抗体复合物（Ag-Ab复合物）刺激产生的超氧阴离子（O₂⁻），以及细胞对抗体致敏红细胞的胞内摄取。然而，当PMN用DFP处理并洗涤以去除抑制剂后，它们又能够展现出产生O₂⁻和吞噬的活性。完整的PMN经[³H]DFP标记后，用 Triton N101溶解，再经十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳显示存在几种不同分子量的[³H]DFP标记蛋白，这些蛋白在用冷DFP预处理细胞后消失。然而，在[³H]DFP存在的情况下，用Ag-Ab复合物刺激经DFP预处理的PMN会导致出现一种分子量为40,000的[³H]DFP标记的膜结合蛋白。通过将溶解的PMN和吞噬体在抗Ig抗体 - 琼脂糖4B上进行亲和层析分离出了这种蛋白。尽管该蛋白的酶学性质尚不清楚，但目前所获得的结果表明它是一种假定的、刺激激活的丝氨酸蛋白酶，参与导致负责呼吸爆发的NADPH氧化酶激活和吞噬体形成的触发事件。
