Signaling mechanisms of inhibition of phospholipase D activation by CHS-111 in formyl peptide-stimulated neutrophils
Signaling mechanisms of inhibition of phospholipase D activation by CHS-111 in formyl peptide-stimulated neutrophils
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DOI：
10.1016/j.bcp.2010.10.007
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发表时间：
2011-01-15
影响因子：
5.8
通讯作者：
Wang, Jih-Pyang
中科院分区：
文献类型：
--
作者：
Chang, Ling-Chu;Huang, Tai-Hung;Wang, Jih-Pyang
选择性磷脂酶D （PLO）抑制剂5-氟-2-吲哚基-去氯氟哌啶（FIPI）抑制O-2（中心点）的产生和细胞迁移，但不抑制甲酰基- met -亮氨酸（fMLP）刺激的大鼠中性粒细胞的脱粒。一种新型苄基吲哚唑类化合物2-苄基-3-（4-羟甲基苯基）吲哚唑（CHS-111）抑制O-2（中心点-）生成和细胞迁移，也降低了fMLP-刺激的PLO活性，但不降低苯酚酯刺激的PLO活性（IC50为3.9 +/- 1.2 μ M）。CHS-111抑制PLD1与adp -核糖基化因子（Art） 6和Ras同源物（Rho） A的相互作用，在fmlp刺激的细胞中减少RhoA的膜募集，而在GTP γ s刺激的无细胞系统中没有。CHS-111降低了gtp结合的RhoA的细胞水平、RhoA相关蛋白激酶1的膜募集和下游肌球蛋白轻链2的磷酸化，减弱了磷脂酰肌醇4-磷酸5激酶（PIP5K）与Arf6的相互作用，而仅轻微抑制人Dbs （DH/PH）蛋白的鸟嘌呤核苷酸交换活性，不影响Arf6与Arf6的结合。CHS-111抑制RhoA与Vav的相互作用、膜结合和Vav的磷酸化。CHS-111对Src家族激酶（SFK）的磷酸化没有影响，但减弱了Vav与Lck、Hck、Fgr和Lyn的相互作用。CHS-111还抑制了PLD1与蛋白激酶C (PKC) α、β I和β II同工酶的相互作用，以及PLD1的磷酸化。这些结果表明，CHS-111对fmlp刺激的PLO活性的抑制是由于通过干扰sfk介导的Vav激活，Arf6与PLD1和PIP5K的相互作用减弱，以及大鼠中性粒细胞中Ca2+依赖性PKC的激活来阻断RhoA激活。(C) 2010爱思唯尔公司版权所有。
A selective phospholipase D (PLO) inhibitor 5-fluoro-2-indolyl des-chlorohalopemide (FIPI) inhibited the O-2(center dot-) generation and cell migration but not degranulation in formyl-Met-Leu-Phe(fMLP)-stimulated rat neutrophils. A novel benzyl indazole compound 2-benzyl-3-(4-hydroxymethylphenyl)indazole (CHS-111), which inhibited O-2(center dot-) generation and cell migration, also reduced the fMLP- but not phorbol ester-stimulated PLO activity (IC50 3.9 +/- 1.2 mu M). CHS-111 inhibited the interaction of PLD1 with ADP-ribosylation factor (Art) 6 and Ras homology (Rho) A, and reduced the membrane recruitment of RhoA in fMLP-stimulated cells but not in GTP gamma S-stimulated cell-free system. CHS-111 reduced the cellular levels of GTP-bound RhoA, membrane recruitment of Rho-associated protein kinase 1 and the downstream myosin light chain 2 phosphorylation, and attenuated the interaction between phosphatidylinositol 4-phosphate 5-kinase (PIP5K) and Arf6, whereas it only slightly inhibited the guanine nucleotide exchange activity of human Dbs (DH/PH) protein and did not affect the arfaptin binding to Arf6. CHS-111 inhibited the interaction of RhoA with Vav, the membrane association and the phosphorylation of Vav. CHS-111 had no effect on the phosphorylation of Src family kinases (SFK) but attenuated the interaction of Vav with Lck, Hck, Fgr and Lyn. CHS-111 also inhibited the interaction of PLD1 with protein kinase C (PKC)alpha, beta I and beta II isoenzymes, and the phosphorylation of PLD1. These results indicate that inhibition of fMLP-stimulated PLO activity by CHS-111 is attributable to the blockade of RhoA activation via the interference with SFK-mediated Vav activation, attenuation of the interaction of Arf6 with PLD1 and PIP5K, and the activation of Ca2+-dependent PKC in rat neutrophils. (C) 2010 Elsevier Inc. All rights reserved.