Anandamide biosynthesis catalyzed by the phosphodiesterase GDE1 and detection of glycerophospho-n-acyl ethanolamine precursors in mouse brain
Anandamide biosynthesis catalyzed by the phosphodiesterase GDE1 and detection of glycerophospho-n-acyl ethanolamine precursors in mouse brain
复制标题
DOI：
10.1074/jbc.m707807200
复制
发表时间：
2008-04-04
影响因子：
4.8
通讯作者：
Cravatt, Benjamin F.
中科院分区：
文献类型：
--
作者：
Simon, Gabriel M.;Cravatt, Benjamin F.
大麻素酰胺（AEA）是大麻素受体的内源性配体，是许多生理过程（包括炎症、疼痛和食欲）的良好表征介质。大麻素及其N-酰基乙醇胺（NAE）同类物的生物合成途径仍然是个谜。以前，我们提出了一种酶促途径，用于生产NAE，包括双-O-脱酰N-酰基磷脂酰乙醇胺（NAPE）的α/β-水解酶4（ABDH 4或Abh 4），以形成甘油磷酸（GP）-NAE，然后通过一个身份不明的磷酸二酯酶将这些中间体转化为NAE。在这里，我们报告的检测和测量的GP-NAEs，包括花生四烯酸酰胺前体甘油磷酸-N-花生四烯酸乙醇胺（GP-NArE），作为内源性成分的小鼠脑组织。抑制磷酸二酯酶介导的GP-NAES体外降解导致这些脂质在脑提取物中显著积累，表明通过该途径的快速内源性通量。此外，我们确定了甘油磷酸二酯酶GDE 1，也称为MIR 16，作为一种广泛表达的膜酶，具有强大的GP-NAE磷酸二酯酶活性。总之，这些数据提供了证据的多步骤途径的生产anandamide在神经系统中的顺序行动的Abh 4和GDE 1。
Anandamide (AEA) is an endogenous ligand of cannabinoid receptors and a well characterized mediator of many physiological processes including inflammation, pain, and appetite. The biosynthetic pathway(s) for anandamide and its N-acyl ethanolamine (NAE) congeners remain enigmatic. Previously, we proposed an enzymatic route for producing NAEs that involves the double-O-deacylation of N-acyl phosphatidylethanolamines (NAPEs) by alpha/beta-hydrolase 4 (ABDH4 or Abh4) to form glycerophospho (GP)-NAEs, followed by conversion of these intermediates to NAEs by an unidentified phosphodiesterase. Here, we report the detection and measurement of GP-NAEs, including the anandamide precursor glycerophospho-N-arachidonoylethanolamine (GP-NArE), as endogenous constituents of mouse brain tissue. Inhibition of the phosphodiesterase-mediated degradation of GP-NAEs ex vivo resulted in a striking accumulation of these lipids in brain extracts, suggesting a rapid endogenous flux through this pathway. Furthermore, we identify the glycerophosphodiesterase GDE1, also known as MIR16, as a broadly expressed membrane enzyme with robust GP-NAE phosphodiesterase activity. Together, these data provide evidence for a multistep pathway for the production of anandamide in the nervous system by the sequential actions of Abh4 and GDE1.