Isolation and identification of the enzyme synthesising Up4A from endothelial cells and isolation, identification, and characterization of further "endothelial-derived vasoconstrictive factors" (EDCF)

从内皮细胞中分离和鉴定合成 Up4A 的酶，并进一步分离、鉴定和表征“内皮源性血管收缩因子”(EDCF)

• 批准号: 30164301
• 负责人: Professor Dr. Markus van der Giet
• 金额: --
• 依托单位: Institut für Molekulare Herz-Kreislauf-Forschung (IMCAR)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2006
• 资助国家: 德国
• 起止时间: 2005-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/30164301?language=en
• 关键词: Isolation; identification; enzyme; synthesising; Up4A

The endothelium not only acts as a mechanical barrier between blood and vessel walls, but is also an endocrine organ with multiple regulatory functions. By the identification of NO and endothelin as vasoregulatory factors our knowledge of endothelial-mediated vascular regulation was revolutionized. In literature there are several indications of further ¿endothelialderived vasoconstrictive factors (EDCF). In our own experiments we isolated uridine adenosine tetraphosphate as a further highly effective EDCF from supernatants of stimulated human endothelial cells. Next the question arose how Up4A is being produced in endothelial cells. First preliminary experiments showed that immobilized endothelial cells contain an enzyme, which is capable of synthesizing Up4A from ADP and UDP. Therefore it is the primary aim of the project to isolate and identify the enzyme synthesizing Up4A. Further experiments showed that human endothelial cells secrete further still unknown EDCFs beyond Up4A, which also belong to the class of nucleotides. Since cultivated endothelial cells secrete only small absolute amounts of these EDCFs, we have not yet been able to isolate and identify these compounds in our preliminary experiments. Therefore, in the second part of the proposal we plan to isolate, identify, quantify, and characterize these still unknown EDCFs. After these compounds have been identified, they will have to be synthesized unless they are commercially available, and the effects on vascular regulation will have to be characterized in detail by those bioassays, which are established in the applicant s group. In a third part of the proposed project we will examine the signal transduction of Up4A and of other, additional EDCFs, which will have been identified in this project, using molecular biology techniques.

内皮细胞不仅是血液和血管壁之间的机械屏障，而且是具有多种调节功能的内分泌器官。通过鉴定NO和内皮素作为血管调节因子，我们对内皮介导的血管调节的认识发生了革命性的变化。在文献中，有几种迹象表明进一步内皮源性血管收缩因子（EDCF）。在我们自己的实验中，我们从刺激的人内皮细胞的上清液中分离出尿苷腺苷四磷酸作为进一步高效的EDCF。接下来的问题是Up4A是如何在内皮细胞中产生的。第一个初步实验表明，固定化的内皮细胞含有一种酶，该酶能够从ADP和UDP合成Up4A。因此，分离和鉴定合成Up4A的酶是本项目的主要目的。进一步的实验表明，人类内皮细胞分泌除了Up4A之外的其他未知EDCF，其也属于核苷酸类。由于培养的内皮细胞仅分泌少量绝对量的这些EDCFs，因此我们在初步实验中还不能分离和鉴定这些化合物。因此，在提案的第二部分中，我们计划分离、鉴定、量化和表征这些仍然未知的EDCF。在这些化合物被鉴定之后，除非它们是商业上可获得的，否则它们将必须被合成，并且对血管调节的作用将必须通过申请人小组建立的那些生物测定来详细表征。在拟议项目的第三部分中，我们将使用分子生物学技术研究Up4A和其他额外EDCF的信号转导，这些EDCF将在本项目中被鉴定。

项目成果

Uridine adenosine tetraphosphate (Up4A) is a strong inductor of smooth muscle cell migration via activation of the P2Y2 receptor and cross-communication to the PDGF receptor.

• DOI: 10.1016/j.bbrc.2011.12.088
• 发表时间: 2012-01
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: A. Wiedon;M. Tölle;Joschika Bastine;M. Schuchardt;Tao-Ming Huang;V. Jankowski;J. Jankowski;W. Zidek;M. van der Giet

P2Y purinoceptors as potential emerging therapeutical target in vascular disease.

• DOI: 10.2174/138161212803582504
• 发表时间: 2012-11
• 期刊: Current pharmaceutical design
• 影响因子: 3.1
• 作者: M. Schuchardt;M. Tölle;M. van der Giet

Highly sensitive, selective and rapid LC-MS method for simultaneous quantification of diadenosine polyphosphates in human plasma.

高灵敏度、选择性和快速 LC-MS 方法同时定量人血浆中的二腺苷多磷酸

• DOI: 10.1016/j.jchromb.2014.05.018
• 发表时间: 2014
• 期刊: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
• 作者: Schulz A;Jankowski V;Zidek W;Jankowski J
• 通讯作者: Jankowski J