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: 7357747
• 负责人: DAVID P ROSE
• 金额: $ 2.67万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7357747
• 关键词: CATALYTIC; MECHANISM; INHIBITION; GOLGI; ALPHA

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 dist

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。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，是潜在的抗转移化合物的靶标。我们主要利用CHESS的数据，通过与中间体和几类抑制剂的络合物的结构来分析酶的催化和抑制的原子基础。最近，我们进行了两次高通量的化学文库筛选，寻找新的酶抑制剂。这导致了10-12个不同亲和力和性质的命中。我们感兴趣的是研究这些如何与结构结合。特别是，一些可能是非竞争性抑制剂，可能占据远离催化中心的位置，这将对未来的抑制剂设计感兴趣。我们想使用同步辐射有两个主要原因。首先，我们可以在几分钟内获得一个数据集，而不是几个小时，这将使我们在短时间内快速检查几个这样的复合体。其次，我们可以从这些晶体中获得几乎原子分辨率(1.2-1.4A)的数据。这一分辨率对于理解构象扭曲和确定精确的原子间距离是至关重要的