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.

蛋白质二硫键异构酶（PDI）催化二硫键的可逆形成和异构化最近的报告表明，PDI 家族的其他成员 ERp57， ERp5 和 ERp72 是否也有助于血栓形成。血栓形成是明显的还是多余的尚不清楚。 ERp57 缺失的小鼠只能通过缺失的特定 PDI 来恢复，这意味着这些酶。在激活支持血小板聚集的 αIIbβ3 血小板整合素方面具有单独的目标。 PDI 家族成员含有 CGHC 活性位点基序，可催化蛋白质构象变化该提案重点关注具有该基序的 PDI 家族的两个新成员：存在于血小板中；ERp46 和 PDI 家族的跨膜成员 TMX3。 ERp57、ERp5 和 ERp72 介导血小板聚集和血栓形成，并参与血小板的转化 αIIbβ3 至其高亲和力状态然而，这些 PDI 调节 αIIbβ3 的实际机制和血小板聚集的每种酶的单独靶点以及它们如何共同发挥作用尚不清楚。为了确定血小板中每种 PDI 的具体功能，我们使用了靶向药物。敲除小鼠方法的具体目的是： 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；表征半胱氨酸/二硫键靶标以及 PDI、ERp57、ERp72 激活 αIIbβ3 的机制， ERp46 和 TMX3 所使用的主要技术是激光诱导的血栓形成损伤模型。确定 PDI 家族多个成员协同工作的实际机制，我们将采用硫醇标记策略与标记硫醇的质谱鉴定这将开始解开谜团。这些酶单独发挥作用的机制，以及它们如何作为网络一起发挥作用。 αIIbβ3 激活的最后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一个方面，并且可以作为其他整合素激活的模型。定义具体机制还可能导致新型抑制剂的出现，这些抑制剂可以双重调节血小板和由于血小板参与多种疾病状态，我们的研究结果可能会有更广泛的范围。对其他疾病状况的基本了解以及可能的治疗方法的影响这些条件。

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）

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

一种新的抗血栓策略：抑制蛋白质二硫键异构酶的C端活性位点。

DOI：
发表时间：
2017-04
期刊：
影响因子：
0
作者：
Wang, L;Essex, D W
通讯作者：
Essex, D W

Thiols in the alphaIIbbeta3 integrin are necessary for platelet aggregation.

αIIbβ3 整合素中的硫醇是血小板聚集所必需的。

DOI：
发表时间：
2008-07
期刊：
影响因子：
6.5
作者：
Manickam, Nagaraj;Sun, Xiuhua;Hakala, Kevin W;Weintraub, Susan T;Essex, David W
通讯作者：
Essex, David W

Platelet-derived ERp57 mediates platelet incorporation into a growing thrombus by regulation of the αIIbβ3 integrin.

血小板衍生的 ERp57 通过调节 αIIbβ3 整合素介导血小板掺入生长中的血栓。

DOI：
10.1182/blood-2013-06-506691
发表时间：
2013-11-21
期刊：
Blood
影响因子：
20.3
作者：
Lu Wang;Yi Wu;Junsong Zhou;S. S. Ahmad;Bulent Mutus;N. Garbi;Günter Hämmerling;Junling Liu
通讯作者：
Junling Liu

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

二硫键异构酶 ERp57 是体内纤维蛋白沉积所必需的。

DOI：
发表时间：
2014-11
期刊：
影响因子：
0
作者：
Zhou, J;Wu, Y;Wang, L;Rauova, L;Hayes, V M;Poncz, M;Essex, D W
通讯作者：
Essex, D W

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

内质网蛋白 46 (ERp46) 在小鼠血小板功能和动脉血栓形成中的新作用。

DOI：
10.1182/blood.2021012055
发表时间：
2021-11-09
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：
D. Essex