ROLE OF TYROSYL PHOSPHORYLATION IN NEUTROPHIL PRIMING BY TUMOR-NECROSIS-FACTOR-ALPHA AND GRANULOCYTE COLONY STIMULATING FACTOR
ROLE OF TYROSYL PHOSPHORYLATION IN NEUTROPHIL PRIMING BY TUMOR-NECROSIS-FACTOR-ALPHA AND GRANULOCYTE COLONY STIMULATING FACTOR
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DOI：
10.1016/0003-9861(92)90469-d
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发表时间：
1992-11-01
影响因子：
3.9
通讯作者：
UTSUMI, K
中科院分区：
文献类型：
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作者：
AKIMARU, K;UTSUMI, T;UTSUMI, K
用抗酪氨酸磷酸化抗体对人肿瘤坏死因子-α（TNF-α）和人粒细胞集落刺激因子（G-CSF）诱导人外周血中性粒细胞（PMN）蛋白酪氨酸残基磷酸化的能力进行了Western印迹分析。TNF-α和G-CSF均增加了各种蛋白质的酪氨酰磷酸化，例如54-、63-、72-、83-、98-、108-和115-kDa蛋白质。配体刺激的酪氨酰磷酸化的115-kDa的蛋白质是时间和浓度依赖性的。当115 kDa的蛋白质被磷酸化，它被回收从膜馏分。酪氨酸激酶（TK）抑制剂染料木黄酮和α-氰基-3-乙氧基-4-羟基-5-苯硫甲基肉桂酰胺（ST 638）可抑制115-kDa蛋白的磷酸化，钙离子和磷脂依赖性蛋白激酶（PKC）抑制剂1-（5-异喹啉磺酰基）甲基哌嗪二盐酸盐（H-7）和星形孢菌素可增强115-kDa蛋白的磷酸化。用TNF-α或G-CSF致敏的PMN，在甲酰甲硫氨酰-亮氨酰苯丙氨酸（FMLP）诱导的超氧阴离子（O-[2]）生成中，也观察到TK抑制剂的类似抑制作用和PKC抑制剂的类似刺激作用。115 kDa蛋白的磷酸化与O β 2的配体依赖性生成平行发生。这些和其他观察结果表明，酪氨酸激酶的底物蛋白，如115 kDa的蛋白质，可能在中性粒细胞的启动机制中发挥关键作用。这是第一份描述酪氨酰磷酸化参与G-CSF和TNF-α致敏中性粒细胞的报告。
The ability of human tumor necrosis factor-α (TNF-α) and human granulocyte colony stimulating factor (G-CSF) to induce phosphorylation of protein tyrosyl residues in human peripheral neutrophils (PMN) was investigated by Western blot analysis with antiphosphotyrosine antibody. Both TNF-α and G-CSF increased the tyrosyl phosphorylation of various proteins, such as species of 54-, 63-, 72-, 83-, 98-, 108-, and 115-kDa proteins. The ligand-stimulated tyrosyl phosphorylation of the 115-kDa protein was time- and concentration-dependent. When the 115-kDa protein was phosphorylated, it was recovered from membrane fractions. The phosphorylation of the 115-kDa protein was inhibited by genistein and α-cyano-3-ethoxy-4-hydroxy-5-phenylthiomethylcinnamamide (ST 638), inhibitors of tyrosine kinase (TK), and was enhanced by 1-(5-isoquinoline-sulfonyl) methyl-piperazine dihydrochloride (H-7) and staurosporine, inhibitors of Ca2+- and phospholipid-dependent protein kinase (PKC). Similar inhibition by the TK inhibitors and stimulation by the PKC inhibitors were also observed with formylmethionyl-leucylphenylalanine (FMLP)-induced superoxide (O⨪2) generation by TNF-α- or G-CSF-primed PMN. Phosphorylation of the 115-kDa protein occurred in parallel with the ligand-dependent generation of O⨪2. These and other observations suggested that substrate proteins for tyrosine kinase, such as the 115-kDa protein, might play critical roles in the mechanism for priming of neutrophils. This is the first report describing that tyrosyl phosphorylation is involved in the priming of neutrophils by G-CSF and TNF-α.