The structural and thermodynamic dissection of the interdomain cooperativity of human ACE

心血管疾病的经济负担是惊人的，据估计，英国每年的成本为500亿英镑，欧盟为2500亿欧元，美国为1750亿美元。人躯体血管紧张素- 1转换酶[ACE，含有两个结构域（N-和C-）]抑制剂被广泛用于治疗心血管疾病，包括高血压、心力衰竭、冠状动脉疾病、纤维化和肾衰竭。然而，当前一代的ACE抑制剂是在20世纪70年代和80年代开发的，由于常见的副作用而受到阻碍。ACE的N-和c -结构域表现出不同的底物特异性。虽然市场上有许多ACE抑制剂可以阻断这两个结构域，但目前还没有一种药物可以选择性地抑制n结构域，从而获得减少心脏、肾脏和肺部纤维化和炎症的优势，而不会伴随阻断c结构域引起的副作用。这强调了确定整个ACE分子结构和设计更安全有效的第二代新型ACE抑制剂复合物的重要性。我们利用x射线晶体学技术成功测定了人类睾丸ACE的晶体结构（相当于体细胞ACE的c结构域）和体细胞ACE的n结构域（具有各种临床重要抑制剂以及新型结构域选择性抑制剂），为真正基于结构设计更好的ACE抑制剂提供了平台。这是对ACE结构细节的重大突破，更重要的是对ACE抑制机制的重大突破。这些重要的结果为更严格的方法铺平了道路，通过基于结构的新型结构域选择性抑制剂的药物设计途径来利用结构域之间的差异。因此，在了解ACE“隐藏”分子特性方面的持续努力为我们的团队提供了一个坚实的平台，以进行我们在提案中概述的新研究。我们提出的实验建立在先前和当前工作的基础上，针对全长ACE的结构研究。该提案的重点是ACE的关键结构-功能研究，解决新的和重要的生物学问题，这将对设计新一代ACE选择性抑制剂结合基础和转化研究产生影响（从长远来看）。最终的长期目标将是设计针对ACE的N端或c端结构域的新化合物，并期望这将提供副作用更小的选择性化合物。

The economic burden of cardiovascular illness is staggering, with estimated annual costs of £50 billion in the UK, Euro250 billion in the EU and $175 billion in the USA.Human somatic angiotensin-I converting enzyme [ACE, which contains two domains (N- and C-)] inhibitors are widely used to treat cardiovascular diseases, including high blood pressure, heart failure, coronary artery disease, fibrosis and kidney failure. However, current-generation ACE inhibitors, which were developed in the 1970's and 1980's, are hampered by common side effects. The N- and C-domains of ACE display different substrate specificities.While there are many ACE inhibitors on the market that block both domains, there are no drugs that selectively inhibit the N-domain and thereby accrue the advantages of reducing fibrosis and inflammation in the heart, kidney and lung, without the concomitant side effects induced by blockade of the C-domain. This underscores the importance of the determination of the molecular structure of entire ACE and the design of 2nd generation of new ACE-inhibitor complex/s that are safer and more effective. Our success in the determination of the crystal structure of human testis ACE (equivalent to the C-domain of somatic ACE) and the N-domain of somatic ACE (with various clinically important inhibitors as well as novel domain selective inhibitors) using the technique X-ray crystallography have provided the platform for true structure-based design of better ACE inhibitors. This is a significant breakthrough in terms of the structural details of ACE and, more importantly, the mechanism of ACE inhibition. These important results pave the way for a more rigorous approach exploiting the differences between the domains through a structure based drug design route of novel domain-selective inhibitors. Thus the sustained effort on understanding the 'hidden' molecular properties of ACE has provided a firm platform for our group for new studies as outlined in our proposal.Our proposed experiments that builds on a body of previous and current work are directed at structural study of the full-length ACE. The proposal has its focus on the crucial structure-function studies of ACE addressing new and important biological questions which will have implications (in the longer term) for the design of new generation of selective inhibitors of ACE combining basic and translational research. The ultimate longer term aim will be the design of new compounds that are specific for the N- or C-terminal domain of ACE with the expectation that this will provide selective compounds with fewer side effects.