Inhibition of monometalated methionine aminopeptidase: Inhibitor discovery and crystallographic analysis
Inhibition of monometalated methionine aminopeptidase: Inhibitor discovery and crystallographic analysis
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DOI：
10.1021/jm700930k
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发表时间：
2007-11-15
影响因子：
7.3
通讯作者：
Ye, Qi-Zhuang
中科院分区：
文献类型：
--
作者：
Huang, Min;Xie, Sheng-Xue;Ye, Qi-Zhuang
甲硫氨酸氨肽酶的活性位点空腔中常见两种二价金属离子，并且至少一种金属离子直接参与催化。尽管二金属化酶有充足的结构和功能信息，但甲硫氨酸氨肽酶在生理条件下可能充当单金属化酶。缺乏有关单金属化酶的结构以及催化和抑制的信息。通过改进高通量筛选条件，我们鉴定了一种对单金属化酶具有特异性的独特抑制剂。动力学表征表明抑制剂与活性位点的第二金属离子之间的结合具有相互排斥性。这通过X射线结构得到证实，并且该抑制剂与第一金属离子配位并占据通常由第二金属离子占据的空间。该抑制剂和其他抑制剂的抑制作用的动力学和结构分析为设计有效的甲硫氨酸氨肽酶抑制剂提供了见解。
Two divalent metal ions are commonly seen in the active-site cavity of methionine aminopeptidase, and at least one of the metal ions is directly involved in catalysis. Although ample structural and functional information is available for dimetalated enzyme, methionine aminopeptidase likely functions as a monometalated enzyme under physiological conditions. Information on structure, as well as catalysis and inhibition, of the monometalated enzyme is lacking. By improving conditions of high-throughput screening, we identified a unique inhibitor with specificity toward the monometalated enzyme. Kinetic characterization indicates a mutual exclusivity in binding between the inhibitor and the second metal ion at the active site. This is confirmed by X-ray structure, and this inhibitor coordinates with the first metal ion and occupies the space normally occupied by the second metal ion. Kinetic and structural analyses of the inhibition by this and other inhibitors provide insight in designing effective inhibitors of methionine aminopeptidase.