Discovery of selective small-molecule activators of a bacterial glycoside hydrolase.
Discovery of selective small-molecule activators of a bacterial glycoside hydrolase.
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DOI：
10.1002/anie.201407081
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发表时间：
2014-12-01
期刊：
影响因子：
--
通讯作者：
Hubbard RE
中科院分区：
文献类型：
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作者：
Darby JF;Landström J;Roth C;He Y;Davies GJ;Hubbard RE
基于片段的方法通常用于发现酶抑制剂作为细胞工具和潜在的治疗剂。然而，关于小分子酶激活剂的发现的报道很少。在此，我们描述了糖苷水解酶（细菌O-GlcNAc水解酶）的小分子活化剂的发现和表征。对市售片段文库进行配体观察的NMR筛选，鉴定出一种酶活化剂，其产生kcat/KM值增加约90%（kcat=催化速率常数; KM=米氏常数）。 该化合物在催化中心附近与酶结合。初始命中的演变导致改进的化合物，其表现为影响KM和Vmax值（Vmax=最大反应速率）的非必需活化剂。 这些化合物似乎可以稳定酶的活性“封闭”形式。这种活化剂可以提供酶抑制剂的正交替代物，用于体内酶活性的扰动，并且还可以用于许多工业过程中的糖苷水解酶活化。
Fragment-based approaches are used routinely to discover enzyme inhibitors as cellular tools and potential therapeutic agents. There have been few reports, however, of the discovery of small-molecule enzyme activators. Herein, we describe the discovery and characterization of small-molecule activators of a glycoside hydrolase (a bacterial O-GlcNAc hydrolase). A ligand-observed NMR screen of a library of commercially available fragments identified an enzyme activator which yielded an approximate 90 % increase in kcat/KM values (kcat=catalytic rate constant; KM=Michaelis constant). This compound binds to the enzyme in close proximity to the catalytic center. Evolution of the initial hits led to improved compounds that behave as nonessential activators effecting both KM and Vmax values (Vmax=maximum rate of reaction). The compounds appear to stabilize an active “closed” form of the enzyme. Such activators could offer an orthogonal alternative to enzyme inhibitors for perturbation of enzyme activity in vivo, and could also be used for glycoside hydrolase activation in many industrial processes.