Fezza Filomena, Oddi Sergio, Di Tommaso Monia, De Simone Chiara, Rapino Cinzia, Pasquariello Nicoletta, Dainese Enrico, Finazzi-Agrò Alessandro, Maccarrone Mauro
European Center for Brain Research/Istituto di Ricovero e Cura a Carattere Scientifico S. Lucia Foundation, Rome, Italy.
J Lipid Res. 2008 Jun;49(6):1216-23. doi: 10.1194/jlr.M700486-JLR200. Epub 2008 Mar 3.
Anandamide (N-arachidonoylethanolamide; AEA) acts as an endogenous agonist of both cannabinoid and vanilloid receptors. During the last two decades, its metabolic pathways and biological activity have been investigated extensively and relatively well characterized. In contrast, at present, the effective nature and mechanism of AEA transport remain controversial and still unsolved issues. Here, we report the characterization of a biotinylated analog of AEA (b-AEA) that has the same lipophilicity of the parent compound. In addition, by means of biochemical assays and fluorescence microscopy, we show that b-AEA is accumulated inside the cells in a way superimposable on that of AEA. Conversely, b-AEA does not interact or interfere with the other components of the endocannabinoid system, such as type-1 and type-2 cannabinoid receptors, vanilloid receptor, AEA synthetase (N-acylphosphatidylethanolamine-hydrolyzing phospholipase D), or AEA hydrolase (fatty acid amide hydrolase). Together, our data suggest that b-AEA could be a very useful probe for visualizing the accumulation and intracellular distribution of this endocannabinoid.
花生四烯酸乙醇胺（N-花生四烯酰乙醇胺；AEA）作为大麻素受体和香草酸受体的内源性激动剂。在过去二十年中，其代谢途径和生物活性已得到广泛研究且特征相对明确。相比之下，目前AEA转运的有效性质和机制仍存在争议，仍是未解决的问题。在此，我们报告了一种与母体化合物具有相同亲脂性的AEA生物素化类似物（b-AEA）的特征。此外，通过生化分析和荧光显微镜观察，我们发现b-AEA在细胞内的积累方式与AEA的积累方式相同。相反，b-AEA不与内源性大麻素系统的其他成分相互作用或干扰，如1型和2型大麻素受体、香草酸受体、AEA合成酶（N-酰基磷脂酰乙醇胺水解磷脂酶D）或AEA水解酶（脂肪酸酰胺水解酶）。总之，我们的数据表明b-AEA可能是一种非常有用的探针，用于可视化这种内源性大麻素的积累和细胞内分布。
