Protective role of the dynamin inhibitor Dynasore against the cholesterol-dependent cytolysin of Trueperella pyogenes.
Protective role of the dynamin inhibitor Dynasore against the cholesterol-dependent cytolysin of Trueperella pyogenes.
复制标题
DOI：
10.1096/fj.14-265207
复制
发表时间：
2015-04
期刊：
影响因子：
--
通讯作者：
Sheldon IM
中科院分区：
文献类型：
--
作者：
Preta G;Lotti V;Cronin JG;Sheldon IM
许多革兰氏阳性细菌的毒力取决于胆固醇依赖性溶细胞素（CDC），它在真核细胞质膜上形成孔。来自化脓性 Trueperella 的脓溶素 (PLO) 为探索细胞对 CDC 的反应提供了独特的机会，因为它不需要硫醇激活。亚溶解浓度的 PLO 刺激原代基质细胞中 MAPK ERK 和 p38 的磷酸化，并通过蛋白质轻链 3B 裂解确定诱导自噬。尽管如此，MAPK 或自噬抑制剂并不影响 PLO 诱导的细胞溶解。然而，10 μM 3-羟基萘-2-羧酸-(3,4-二羟基亚苯亚甲基)-酰肼 (Dynasore)（一种动力鸟苷 5'-三磷酸酶抑制剂）可保护基质细胞免受 PLO 诱导的细胞溶解作用（如下测定） 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四唑测定（85 ± 17% 与 50 ± 9% 细胞活力）、测量细胞外 ATP 和动力学测定。这是一种通用机制，因为 Dynasore 还可以保护 HeLa 细胞免受链球菌溶血素 O 的侵害。此外，这种作用是可逆的，基质细胞对 PLO 的敏感性在 Dynasore 去除后 30 分钟内恢复。 Dynasore 的保护作用不是由动力抑制、ERK 磷酸化诱导或 Dynasore 与 PLO 结合赋予的。相反，Dynasore 降低了细胞胆固醇并破坏了质膜脂筏，类似于阳性对照甲基-β-环糊精。 Dynasore 是一种易于处理的工具，可探索真核细胞中胆固醇稳态的复杂性，并制定对抗 CDC 的策略。—Preta, G.、Lotti, V.、Cronin, J. G. 和 Sheldon, I. M. 动力抑制剂 Dynasore 对化脓性 Trueperella 的胆固醇依赖性溶细胞素的保护作用。
The virulence of many Gram-positive bacteria depends on cholesterol-dependent cytolysins (CDCs), which form pores in eukaryotic cell plasma membranes. Pyolysin (PLO) from Trueperella pyogenes provided a unique opportunity to explore cellular responses to CDCs because it does not require thiol activation. Sublytic concentrations of PLO stimulated phosphorylation of MAPK ERK and p38 in primary stromal cells, and induced autophagy as determined by protein light-chain 3B cleavage. Although, inhibitors of MAPK or autophagy did not affect PLO-induced cytolysis. However, 10 μM 3-hydroxynaphthalene-2-carboxylic acid-(3,4-dihydroxybenzylidene)-hydrazide (Dynasore), a dynamin guanosine 5′-triphosphatase inhibitor, protected stromal cells against PLO-induced cytolysis as determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (85 ± 17% versus 50 ± 9% cell viability), measuring extracellular ATP, and kinetic assays. This was a generalized mechanism because Dynasore also protected HeLa cells against streptolysin O. Furthermore, the effect was reversible, with stromal cell sensitivity to PLO restored within 30 minutes of Dynasore removal. The protective effect of Dynasore was not conferred by dynamin inhibition, induction of ERK phosphorylation, or Dynasore binding to PLO. Rather, Dynasore reduced cellular cholesterol and disrupted plasma membrane lipid rafts, similar to positive control methyl-β-cyclodextrin. Dynasore is a tractable tool to explore the complexity of cholesterol homeostasis in eukaryotic cells and to develop strategies to counter CDCs.—Preta, G., Lotti, V., Cronin, J. G., and Sheldon, I. M. Protective role of the dynamin inhibitor Dynasore against the cholesterol-dependent cytolysin of Trueperella pyogenes.
登录
查看更多内容
Dynasore是一种动力蛋白抑制剂，可抑制克鲁齐锥虫进入腹膜巨噬细胞。
影响因子：
3.7
作者：
Barrias ES;Reignault LC;De Souza W;Carvalho TM
通讯作者：
Carvalho TM
影响因子：
3.6
作者：
Cronin JG;Turner ML;Goetze L;Bryant CE;Sheldon IM
通讯作者：
Sheldon IM
影响因子：
3.2
作者：
Billington, SJ;Jost, BH;Songer, JG
通讯作者：
Songer, JG
影响因子：
3.7
作者：
Girard E;Paul JL;Fournier N;Beaune P;Johannes L;Lamaze C;Védie B
通讯作者：
Védie B
影响因子：
3.2
作者：
Gelber, Shari E.;Aguilar, Jorge L.;Ratner, Adam J.
通讯作者：
Ratner, Adam J.