CATALYTIC MECHANISM AND INHIBITION OF GOLGI ALPHA-MANNOSIDASE II

高尔基体α-甘露糖苷酶II的催化机制和抑制作用

• 批准号: 7721296
• 负责人: DAVID ROSE
• 金额: $ 4.06万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721296
• 关键词: Affinity; Alpha-mannosidase; Binding; Catalysis; Class; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Data Set; Distant; Enzyme Inhibition; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Family; Funding; Future; Golgi Apparatus; Grant; Hour; Hydrolase; Institution; Property; Research; Research Personnel; Resolution; Resources; Site; Source; Structure; Time; United States National Institutes of Health; base; design; glycosylation; high throughput screening; inhibitor/antagonist; interest; mannosyl-oligosaccharide 1,3-1,6-alpha-mannosidase; novel; small molecule libraries; synchrotron radiation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We previously determined the structure of the glycosylation enzyme alpha-mannosidase II, a glycosyl hydrolase of the Family 38 family targetted by potential anti-metastatic compounds. Largely with data from CHESS, we have been analyzing the atomic basis of catalysis and inhibition of the enzyme through structures of complexes with intermediates and several classes of inhibitors. Recently, we performed two high-throughput screens of chemical libraries looking for novel inhibitors of the enzyme. This resulted in 10-12 hits of varying affinity and properties. We are interested in examining how these bind to the structure. In particular, some may be non-competitive inhibitors and may occupy sites distant from the catalytic centre, which will be of interest for future inhibitor design. We would like to use synchrotron radiation for two promary reasons. Firstly, we can get a data set in minutes, rather than hours, which will let us quickly examine several of these complexes in a short time. Secondly, we can get almost atomic resolution (1.2-1.4A) data from these crystals at CHESS. This resolution can be critical in understanding conformational distortions and determining precise interatomic distances.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们之前确定了糖基化酶 α-甘露糖苷酶 II 的结构，这是一种 38 家族的糖基水解酶，是潜在抗转移化合物的靶标。我们主要利用 CHESS 的数据，通过中间体和几类抑制剂的复合物结构来分析酶催化和抑制的原子基础。 最近，我们对化学库进行了两次高通量筛选，寻找新型酶抑制剂。这导致了 10-12 次具有不同亲和力和特性的命中。我们有兴趣研究它们如何与结构结合。特别是，一些可能是非竞争性抑制剂，并且可能占据远离催化中心的位点，这对于未来的抑制剂设计将是有意义的。 我们希望使用同步加速器辐射有两个主要原因。首先，我们可以在几分钟而不是几小时内获得数据集，这将使我们能够在短时间内快速检查其中的几个复合体。其次，我们可以在 CHESS 中从这些晶体中获得几乎原子分辨率（1.2-1.4A）的数据。这种分辨率对于理解构象扭曲和确定精确的原子间距离至关重要。