Binding and inactivation mechanism of a humanized fatty acid amide hydrolase by alpha-ketoheterocycle inhibitors revealed from cocrystal structures.
Binding and inactivation mechanism of a humanized fatty acid amide hydrolase by alpha-ketoheterocycle inhibitors revealed from cocrystal structures.
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α-酮二型抑制剂的人性化脂肪酸酰胺水解酶的结合和灭活机制揭示了共晶结构。
DOI：
10.1021/ja902694n
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发表时间：
2009-08-05
影响因子：
15
通讯作者：
Stevens, Raymond C.
中科院分区：
文献类型：
--
作者：
Mileni, Mauro;Garfunkle, Joie;DeMartino, Jessica K.;Cravatt, Benjamin F.;Boger, Dale L.;Stevens, Raymond C.
公开了与脂肪酸酰胺水解酶（FAAH）结合的两种异构体α-酮恶唑抑制剂（1（OL-135）和2）的共晶X射线结构，所述脂肪酸酰胺水解酶是内源性大麻素信号传导的关键酶调节剂。活性位点催化Ser 241与抑制剂的亲电羰基共价结合，提供与抑制剂结合的FAAH的第一结构作为去质子化的半缩酮，模拟酶促四面体中间体。这项工作还提供了一个详细的氧阴离子孔和一个特殊的“在行动”描绘不寻常的丝氨酸-丝氨酸-赖氨酸催化三元组。这些结构捕捉到的第一张照片的抑制剂，跨越到细胞溶质端口的活性位点提供新的见解，有助于解释FAAH与底物离去基团的相互作用，以及它们在调节抑制剂的效力和选择性的作用。活化中心杂环的作用被明确定义，并与丝氨酸蛋白酶的先前应用中观察到的作用区分开，调和了这类抑制剂所特有的连接取代基的大电子效应与FAAH。额外的引人注目的活性位点的灵活性被认为是结合后的抑制剂，提供了一个现在定义明确的膜通道的存在与空间上独立的酰基链结合口袋的消失的见解。最后，OL-135（1）及其异构体2的结构比较表明，尽管中心活化杂环的方向相反，但它们与FAAH的结合方式相同，这表明末端2-吡啶基取代基和酰基链苯基提供了关键的锚定相互作用，并证实了活化恶唑的独特作用。
The co-crystal X-ray structures of two isomeric α-ketooxazole inhibitors (1 (OL-135) and 2) bound to fatty acid amide hydrolase (FAAH), a key enzymatic regulator of endocannabinoid signaling, are disclosed. The active site catalytic Ser241 is covalently bound to the inhibitors’ electrophilic carbonyl groups, providing the first structures of FAAH bound to an inhibitor as a deprotonated hemiketal mimicking the enzymatic tetrahedral intermediate. The work also offers a detailed view of the oxyanion hole and an exceptional “in-action” depiction of the unusual Ser-Ser-Lys catalytic triad. These structures capture the first picture of inhibitors that span the active site into the cytosolic port providing new insights that help to explain FAAH’s interaction with substrate leaving groups and their role in modulating inhibitor potency and selectivity. The role for the activating central heterocycle is clearly defined and distinguished from that observed in prior applications with serine proteases, reconciling the large electronic effect of attached substituents found unique to this class of inhibitors with FAAH. Additional striking active site flexibility is seen upon binding of the inhibitors, providing insights into the existence of a now well-defined membrane access channel with the disappearance of a spatially independent acyl chain-binding pocket. Finally, comparison of the structures of OL-135 (1) and its isomer 2 indicates that they bind identically to FAAH, albeit with reversed orientations of the central activating heterocycle, revealing that the terminal 2-pyridyl substituent and the acyl chain phenyl group provide key anchoring interactions and confirming the distinguishing role of the activating oxazole.
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影响因子：
1.7
作者：
Duan, GL;Smith, VH;Weaver, DF
通讯作者：
Weaver, DF
影响因子：
15
作者：
Guimaraes, CRW;Boger, DL;Jorgensen, WL
通讯作者：
Jorgensen, WL
影响因子：
64.8
作者：
Calignano, A;La Rana, G;Piomelli, D
通讯作者：
Piomelli, D
影响因子：
56.9
作者：
CRAVATT, BF;PROSPEROGARCIA, O;LERNER, RA
通讯作者：
LERNER, RA
影响因子：
7.3
作者：
Chang, Leon;Luo, Lin;Webb, Michael
通讯作者：
Webb, Michael