Anandamide amidohydrolase activity, released in the medium by Tetrahymena pyriformis. Identification and partial characterization

FEBS Letters

Fasia, Lambrini¹  Karava, Vivi¹  Siafaka-Kapadai, Athanassia¹ 

[1] Department of Chemistry (Biochemistry), University of Athens, Panepistimioupolis, 15771 Athens, Greece

关键词: Anandamide metabolism;    Fatty acid amidohydrolase;    Secreted enzyme;    Tetrahymena pyriformis;   

DOI  :  10.1016/S0014-5793(01)03095-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Anandamide, an endogenous cannabinoid receptor ligand, was rapidly metabolized by Tetrahymena pyriformis in vivo. Metabolic products were mainly phospholipids as well as neutral lipids, including small amounts of free arachidonic acid. Anandamide amidohydrolase activity was detected in the culture medium by the release of [³H]arachidonic acid from [³H]anandamide, in a time- and concentration-dependent manner. Kinetic experiments demonstrated that the released enzyme had an apparent K _m of 3.7 μM and V _max 278 pmol/min/mg protein. Amidohydrolase activity was maximal at pH 9–10, was abolished by phenylmethylsulfonyl fluoride and was Ca²⁺- and Mg²⁺-independent. Thus, T. pyriformis is capable of hydrolyzing anandamide in vivo and releasing amidohydrolase activity.

【 授权许可】

Unknown