Protein disulfide isomerase acts as an injury response signal that enhances fibrin generation via tissue factor activation
Protein disulfide isomerase acts as an injury response signal that enhances fibrin generation via tissue factor activation
复制标题
DOI：
10.1172/jci32376
复制
发表时间：
2008-03-01
影响因子：
15.9
通讯作者：
Engelmann, Bernd
中科院分区：
文献类型：
--
作者：
Reinhardt, Christoph;von Bruehl, Marie-Luise;Engelmann, Bernd
启动蛋白组织因子(TF)的激活可能是凝血过程中的关键步骤，从而导致纤维蛋白的形成。诱导转铁蛋白激活的刺激物在很大程度上是未知的。在此，我们发现氧化还原酶蛋白二硫键异构酶(PDI)在体内血栓形成过程中直接促进依赖于TF的纤维蛋白的产生。小鼠颈动脉内皮剥脱后，PDI在损伤部位从黏附的血小板和破坏的管壁细胞中释放出来。抑制PDI减少了体内不同血栓形成小鼠模型中TF触发的纤维蛋白形成，正如活体荧光显微镜所确定的那样。PDI输注增加--在减少血小板粘附性的条件下，PDI抑制减少--损伤部位的纤维蛋白生成，表明PDI可以直接启动凝血。在体外，人血小板分泌的PDI有助于激活微泡(微粒)上的隐形转铁蛋白。质谱分析表明，Tf胞外半胱氨酸209的一部分是由谷胱甘肽组成的。混合二硫键的形成有助于保持Tf处于低官能度状态。我们认为，还原的PDI通过将混合二硫键和游离的硫醇异构化为分子内二硫键来激活Tf。我们的发现表明，二硫键异构酶可以作为损伤反应信号，在血管损伤后触发纤维蛋白形成的激活。
The activation of initiator protein tissue factor (TF) is likely to be a crucial step in the blood coagulation process, which leads to fibrin formation. The stimuli responsible for inducing TF activation are largely undefined. Here we show that the oxidoreductase protein disulfide isomerase (PDI) directly promotes TF-dependent fibrin production during thrombus formation in vivo. After endothelial denudation of mouse carotid arteries, PDI was released at the injury site from adherent platelets and disrupted vessel wall cells. Inhibition of PDI decreased TF-triggered fibrin formation in different in vivo murine models of thrombus formation, as determined by intravital fluorescence microscopy. PDI infusion increased - and, under conditions of decreased platelet adhesion, PDI inhibition reduced - fibrin generation at the injury site, indicating that PDI can directly initiate blood coagulation. In vitro, human platelet-secreted PDI contributed to the activation of cryptic TF on microvesicles (microparticles). Mass spectrometry analyses indicated that part of the extracellular cysteine 209 of TF was constitutively glutathionylated. Mixed disulfide formation contributed to maintaining TF in a state of low functionality. We propose that reduced PDI activates TF by isomerization of a mixed disulfide and a free thiol to an intramolecular disulfide. Our findings suggest that disulfide isomerases can act as injury response signals that trigger the activation of fibrin formation following vessel injury.