Inactivation of N-acyl phosphatidylethanolamine phospholipase D reveals multiple mechanisms for the biosynthesis of endocannabinoids
Inactivation of N-acyl phosphatidylethanolamine phospholipase D reveals multiple mechanisms for the biosynthesis of endocannabinoids
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DOI：
10.1021/bi060163l
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发表时间：
2006-04-18
期刊：
影响因子：
2.9
通讯作者：
Cravatt, BF
中科院分区：
文献类型：
--
作者：
Leung, D;Saghatelian, A;Cravatt, BF
N-酰基乙醇胺（NAE）构成了包括内源性大麻素anandamide在内的大量且多样的一类信号传导脂质。与其他脂质递质一样，NAE被认为是生物合成和按需降解的，而不是在信号传导之前储存在囊泡中。因此，鉴定参与NAE代谢的酶对于完全理解这种脂质信号传导系统并控制其以获得潜在的治疗效果至关重要。最近，N-酰基磷脂酰乙醇胺磷脂酶D（NAPE PLD）被确定为参与NAE生物合成的候选酶。在这里，我们描述了在NAPE-PLD基因中具有靶向破坏的小鼠[NAPE-PLD（-/-）小鼠]的产生和表征。NAPE-PLD（-/-）小鼠的脑组织显示，NAPE向带有饱和和多不饱和N-酰基链的NAE的钙依赖性转化减少了5倍以上。然而，只有前一组NAE在NAPE-PLD（-/-）脑中的水平降低，并且这些降低对于具有非常长的酰基链（：00）的NAE最为显著。进一步的研究在接受多种NAPE作为底物的NAPE-PLD（-/-）小鼠的脑中鉴定了钙非依赖性PLD活性，包括花生四烯酸酰胺前体C20：4 NAPE。不同的酶途径的生物合成的长链饱和和多不饱和NAE的照明建议的策略，以控制这些脂质的特定子集的活动，而不全局影响作为一个整体的NAE家族的功能。
N-Acyl ethanolamines (NAEs) constitute a large and diverse class of signaling lipids that includes the endogenous cannabinoid anandamide. Like other lipid transmitters, NAEs are thought to be biosynthesized and degraded on-demand rather than being stored in vesicles prior to signaling. The identification of enzymes involved in NAE metabolism is therefore imperative to achieve a complete understanding of this lipid signaling system and control it for potential therapeutic gain. Recently, an N-acyl phosphatidylethanolamine phospholipase D (NAPE-PLD) was identified as a candidate enzyme involved in the biosynthesis of NAEs. Here, we describe the generation and characterization of mice with a targeted disruption in the NAPE-PLD gene [NAPE-PLD(-/-) mice]. Brain tissue from NAPE-PLD(-/-) mice showed more than a 5-fold reduction in the calcium-dependent conversion of NAPEs to NAEs bearing both saturated and polyunsaturated N-acyl chains. However, only the former group of NAEs was decreased in level in NAPE-PLD(-/-) brains, and these reductions were most dramatic for NAEs bearing very long acyl chains ( :00). Further studies identified a calcium-independent PLD activity in brains from NAPE-PLD(-/-) mice that accepted multiple NAPEs as substrates, including the anandamide precursor C20:4 NAPE. The illumination of distinct enzymatic pathways for the biosynthesis of long chain saturated and polyunsaturated NAEs suggests a strategy to control the activity of specific subsets of these lipids without globally affecting the function of the NAE family as a whole.