Infection Signaling Network Research Center, Research Institute of Medical Sciences, Department of Physiology, School of Medicine, Chungnam National University, Daejeon 301-131, Korea.
Korean J Physiol Pharmacol. 2012 Jun;16(3):199-204. doi: 10.4196/kjpp.2012.16.3.199. Epub 2012 Jun 26.
We evaluated the role of Tat-mediated p66shc transduction on the activation of endothelial nitric oxide synthase in cultured mouse endothelial cells. To construct the Tat-p66shc fusion protein, human full length p66shc cDNA was fused with the Tat-protein transduction domain. Transduction of TAT-p66shc showed a concentration- and time-dependent manner in endothelial cells. Tat-mediated p66shc transduction showed increased hydrogen peroxide and superoxide production, compared with Tat-p66shc (S/A), serine 36 residue mutant of p66shc. Tat-mediated p66shc transduction decreased endothelial nitric oxide synthase phosphorylation in endothelial cells. Furthermore, Tat-mediated p66shc transduction augmented TNF-α-induced p38 MAPK phosphorylation in endothelial cells. These results suggest that Tat-mediated p66shc transduction efficiently inhibited endothelial nitric oxide synthase phosphorylation in endothelial cells.
我们评估了 Tat 介导的 p66shc 转导在培养的小鼠内皮细胞中内皮型一氧化氮合酶激活中的作用。为了构建 Tat-p66shc 融合蛋白，将人全长 p66shc cDNA 与 Tat 蛋白转导结构域融合。Tat-p66shc 在血管内皮细胞中的转导呈浓度和时间依赖性。与 Tat-p66shc（S/A），p66shc 的丝氨酸 36 残基突变体相比，Tat 介导的 p66shc 转导显示出增加的过氧化氢和超氧化物产生。Tat 介导的 p66shc 转导降低了内皮细胞中内皮型一氧化氮合酶的磷酸化。此外，Tat 介导的 p66shc 转导增强了 TNF-α 诱导的内皮细胞中 p38 MAPK 磷酸化。这些结果表明，Tat 介导的 p66shc 转导有效地抑制了内皮细胞中内皮型一氧化氮合酶的磷酸化。
