The putative endocannabinoid transport blocker LY2183240 is a potent inhibitor of FAAH and several other brain serine hydrolases
The putative endocannabinoid transport blocker LY2183240 is a potent inhibitor of FAAH and several other brain serine hydrolases
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DOI：
10.1021/ja062999h
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发表时间：
2006-08-02
影响因子：
15
通讯作者：
Cravatt, Benjamin F.
中科院分区：
文献类型：
--
作者：
Alexander, Jessica P.;Cravatt, Benjamin F.
脂质递质如何在细胞内和细胞之间移动以传递信号仍然是一个重要的、在很大程度上尚未回答的问题。整合膜转运蛋白、可溶性脂质结合蛋白和代谢酶都被认为可以协同调节体内的脂质信号动力学。要分配每一类蛋白质的相对贡献，需要选择性的药理试剂来干扰它们的单独功能。最近，LY2183240，一种可能的内源性大麻素转运体的杂环尿素抑制剂，被证明在体内可以干扰细胞对脂质内源性大麻酰胺的摄取，并促进镇痛。在这里，我们证明了LY2183240是EC降解酶脂肪酸酰胺水解酶(FAAH)的有效共价抑制剂。LY2183240通过酶的丝氨酸亲核试剂的氨甲基化来灭活FAAH。使用基于活性的蛋白质组探针的更多全球筛选发现了几个额外的丝氨酸水解酶，这些酶也被LY2183240抑制。这些结果表明，LY2183240对ANANDAME摄取的抑制可能主要是由于FAAH的失活，进一步证明了该酶作为代谢驱动力促进了ANANDAME向细胞内的扩散。更广泛地说，LY2183240的蛋白质组范围的靶标混杂将杂环尿素指定为一种具有潜在过度蛋白质反应性的化学类型，用于药物设计。
How lipid transmitters move within and between cells to communicate signals remains an important and largely unanswered question. Integral membrane transporters, soluble lipid-binding proteins, and metabolic enzymes have all been proposed to collaboratively regulate lipid signaling dynamics in vivo. Assignment of the relative contributions made by each of these classes of proteins requires selective pharmacological agents to perturb their individual functions. Recently, LY2183240, a heterocyclic urea inhibitor of the putative endocannabinoid (EC) transporter, was shown to disrupt the cellular uptake of the lipid EC anandamide and promote analgesia in vivo. Here, we show that LY2183240 is a potent, covalent inhibitor of the EC-degrading enzyme fatty acid amide hydrolase (FAAH). LY2183240 inactivates FAAH by carbamylation of the enzyme's serine nucleophile. More global screens using activity-based proteomic probes identified several additional serine hydrolases that are also inhibited by LY2183240. These results indicate that the blockade of anandamide uptake observed with LY2183240 may be due primarily to the inactivation of FAAH, providing further evidence that this enzyme serves as a metabolic driving force that promotes the diffusion of anandamide into cells. More generally, the proteome-wide target promiscuity of LY2183240 designates the heterocyclic urea as a chemotype with potentially excessive protein reactivity for drug design.