Functional Role of Protein Disulfide Isomerase Isoforms in Platelets

血小板中蛋白质二硫键异构酶亚型的功能作用

基本信息

批准号：
10228655
负责人：
DAVID W ESSEX
金额：
$ 61.35万
依托单位：
TEMPLE UNIV OF THE COMMONWEALTH
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-06-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10228655
关键词：
Active Sites Affinity Biological Blood Cells Blood Platelets Blood coagulation Cardiovascular Diseases Cellular biology Cleaved cell Clot retraction Coagulation Process Cysteine Defect Deposition Disease Disulfides ERp57 Enzymes Event Family Family member Fibrin Glutathione Disulfide Hemorrhage Human Individual Injury Integral Membrane Protein Integrins Knockout Mice Label Laboratories Lasers Lead Literature Mass Spectrum Analysis Mediating Modeling Molecular Conformation Morbidity - disease rate Mus Myocardial Infarction Oxidation-Reduction Platelet Activation Platelet aggregation Prevention Protein Disulfide Isomerase Protein Isoforms Proteins Receptor Activation Receptor Cell Regulation Reporting Role Signal Transduction Source Stroke Sulfhydryl Compounds Surface Techniques Testing Therapeutic Thrombosis Thrombus United States Work base conditional knockout disulfide bond endoplasmic reticulum glycoprotein p72 extracellular inhibitor/antagonist insight member mortality novel platelet function response

项目摘要

Protein disulfide isomerase (PDI) catalyzes the reversible formation and isomerization of disulfide bonds in proteins, and supports thrombosis. Recent reports indicate that other members of the PDI family, ERp57, ERp5, and ERp72 also contribute to thrombosis. Whether the roles of PDI, ERp57, ERp5 and ERp72 in thrombosis were distinct or redundant was unclear. We showed the aggregation defect in ERp72, PDI or ERp57-null mice was only recovered by the specific PDI that was missing. This implies that these enzymes have individual targets in the activation of the αIIbβ3 platelet integrin that supports platelet aggregation. These PDI family members contain the CGHC active-site motif that catalyzes conformational changes in proteins involved in thrombosis. This proposal focuses on two novel members of the PDI family with this motif that are found in platelets; ERp46, and a transmembrane member of the PDI family, TMX3. We now know that PDI, ERp57, ERp5 and ERp72 mediate platelet aggregation and thrombosis, and are involved in conversion of αIIbβ3 to its high affinity state. However, the actual mechanisms by which these PDIs regulate αIIbβ3 and platelet aggregation are unknown. The individual targets of each enzyme and how they function together remains an enigma. To determine the specific function of each PDI in platelets we have used a targeted knockout mice approach. The specific aims are to: 1. Characterize the role of ERp46 in thrombus formation and platelet function; 2. Characterize the role of TMX3 in thrombus formation and platelet function; and 3. Characterize the cysteine/disulfide targets and mechanism of activation of αIIbβ3 by PDI, ERp57, ERp72, ERp46 and TMX3. A principal technique used will be the laser-induced injury model of thrombosis. To determine the actual mechanisms by which multiple members of the PDI family work together we will employ a thiol labeling strategy with mass spectrometry identification of the labeled thiols. This will begin to unravel the mechanisms by which these enzymes work individually, and how they work together as a network. Elucidation of the extracellular redox network required for the final steps in the activation of αIIbβ3 represents a significant aspect of platelet function and thrombus formation, and can be a model for activation of other integrins. Defining the specific mechanisms could also lead to novel types of inhibitors that dually regulate platelets and coagulation. Since platelets are involved in a variety of disease states, our findings will likely have broader implications for basic understanding of other disease conditions, and possible therapeutic approaches for these conditions.

蛋白质二硫化物异构酶（PDI）催化二硫键的可逆形成和异构化蛋白质，并支持血栓形成。最近的报告表明，PDI家族的其他成员ERP57， ERP5和ERP72也有助于血栓形成。 PDI，ERP57，ERP5和ERP72的角色是否血栓形成是不同的或冗余的。我们在ERP72，PDI或 ERP57-NULL小鼠仅被缺少的特定PDI回收。这意味着这些酶具有支持血小板聚集的αIIBβ3血小板整合素的激活中的单个靶标。这些 PDI家族成员包含CGHC主动位点，该主题催化蛋白质的构象变化参与血栓形成。该提案以这个主题为重点是PDI家族的两个新颖成员在血小板中发现； ERP46，以及PDI家族的跨膜成员TMX3。我们现在知道PDI， ERP57，ERP5和ERP72介导血小板聚集和血栓形成，并参与转化 αIIBβ3至其高亲和力状态。但是，这些PDI调节αIIBβ3和的实际机制血小板聚集尚不清楚。每个酶的个别靶标及其如何共同发挥作用仍然是一个谜。为了确定血小板中每个PDI的特定功能，我们使用了一个目标敲除小鼠接近。具体目的是：1。表征ERP46在血栓形成中的作用和血小板功能； 2。表征TMX3在血栓形成和血小板功能中的作用；和3。表征半胱氨酸/二硫化物靶标和PDI，ERP57，ERP72，ERP72，ERP72的激活机理 ERP46和TMX3。使用的主要技术将是激光诱导的血栓形成模型。到确定PDI家族多个成员一起工作的实际机制，我们将采用一个具有标记硫醇的质谱鉴定的硫醇标记策略。这将开始解开这些酶可以单独起作用的机制，以及它们如何作为网络一起工作。阐明激活αIIBβ3所需的最终步骤所需的细胞外氧化还原网络代表了重要的血小板功能和血栓形成的方面，可以成为激活其他整合素的模型。定义特定机制还可能导致新型抑制剂，这些抑制剂双重调节血小板和凝血。由于血小板参与了各种疾病状态，因此我们的发现可能会更广泛对其他疾病疾病的基本理解以及可能的治疗方法的意义这些条件。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Multiple protein disulfide isomerases support thrombosis.

DOI：
10.1097/moh.0000000000000449
发表时间：
2018-09
期刊：
Current opinion in hematology
影响因子：
3.2
作者：
Essex DW;Wu Y
通讯作者：
Wu Y

A new antithrombotic strategy: inhibition of the C-terminal active site of protein disulfide isomerase.

DOI：
10.1111/jth.13634
发表时间：
2017-04
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
0
作者：
Wang L;Essex DW
通讯作者：
Essex DW

The disulfide isomerase ERp57 is required for fibrin deposition in vivo.

DOI：
10.1111/jth.12709
发表时间：
2014-11
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
0
作者：
Zhou J;Wu Y;Wang L;Rauova L;Hayes VM;Poncz M;Essex DW
通讯作者：
Essex DW

A novel role for endoplasmic reticulum protein 46 (ERp46) in platelet function and arterial thrombosis in mice.

DOI：
10.1182/blood.2021012055
发表时间：
2021-11
期刊：
Blood
影响因子：
20.3
作者：
Junsong Zhou;Yi Wu;L. Rauova;Gavin Koma;Lu Wang;M. Poncz;Hong Li;Tong Liu;K. Fong;J. Bennett;S. Kunapuli;D. Essex
通讯作者：
Junsong Zhou;Yi Wu;L. Rauova;Gavin Koma;Lu Wang;M. Poncz;Hong Li;Tong Liu;K. Fong;J. Bennett;S. Kunapuli;D. Essex

Vascular thiol isomerases in thrombosis: The yin and yang.