Protein kinase Cα-RhoA cross-talk in CCL2-induced alterations in brain endothelial permeability

被引:149
作者
Stamatovic, SM
Dimitrijevic, OB
Keep, RF
Andjelkovic, AV
机构
[1] Univ Michigan, Sch Med, Dept Neurosurg, Ann Arbor, MI 48109 USA
[2] Univ Michigan, Sch Med, Dept Pathol, Ann Arbor, MI 48109 USA
[3] Univ Michigan, Sch Med, Dept Mol & Integrat Physiol, Ann Arbor, MI 48109 USA
关键词
D O I
10.1074/jbc.M513122200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Monocyte chemoattractant protein- 1 ( MCP- 1 or CCL2) regulates blood- brain barrier permeability by inducing morphological and biochemical alterations in the tight junction ( TJ) complex between brain endothelial cells. The present study used cultured brain endothelial cells to examine the signaling networks involved in the redistribution of TJ proteins ( occludin, ZO- 1, ZO- 2, claudin-5) by CCL2. The CCL2- induced alterations in the brain endothelial barrier were associated with de novo Ser/ Thr phosphorylation of occludin, ZO- 1, ZO- 2, and claudin- 5. The phosphorylated TJ proteins were redistributed/ localized in Triton X- 100- soluble as well as Triton X- 100- insoluble cell fractions. Two protein kinase C ( PKC) isoforms, PKC alpha and PKC zeta, had a significant impact on this event. Inhibition of their activity using dominant negative mutants PKC alpha- DN and PKC zeta- DN diminished CCL2 effects on brain endothelial permeability. Previous data indicate that Rho/ Rho kinase signaling is involved in CCL2 regulation of brain endothelial permeability. The interactions between the PKC and Rho/ Rho kinase pathways were therefore examined. Rho, PKC alpha, and PKC zeta activities were knocked down using dominant negative mutants ( T17Rho, PKC alpha- DN, and PKC zeta- DN, respectively). PKC alpha and Rho, but not PKC zeta and Rho, interacted at the level of Rho, with PKC alpha being a downstream target for Rho. Double transfection experiments using dominant negative mutants confirmed that this interaction is critical for CCL2- induced redistribution of TJ proteins. Collectively these data suggest for the first time that CCL2 induces brain endothelial hyperpermeability via Rho/ PKC alpha signal pathway interactions.
引用
收藏
页码:8379 / 8388
页数:10
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