| Mammalian cells stably overexpressing N-acylphosphatidylethanolamine-hydrolysing phospholipase D exhibit significantly decreased levels of N-acylphosphatidylethanolamines. | |
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MedLine Citation:
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PMID: 15760304 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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In animal tissues, NAEs (N-acylethanolamines), including N-arachidonoylethanolamine (anandamide), are primarily formed from their corresponding NAPEs (N-acylphosphatidylethanolamines) by a phosphodiesterase of the PLD (phospholipase D) type (NAPE-PLD). Recently, we cloned cDNAs of NAPE-PLD from mouse, rat and human [Okamoto, Morishita, Tsuboi, Tonai and Ueda (2004) J. Biol. Chem. 279, 5298-5305]. However, it remained unclear whether NAPE-PLD acts on endogenous NAPEs contained in the membrane of living cells. To address this question, we stably transfected two mammalian cell lines (HEK-293 and CHO-K1) with mouse NAPE-PLD cDNA, and investigated the endogenous levels and compositions of NAPEs and NAEs in these cells, compared with mock-transfected cells, with the aid of GC-MS. The overexpression of NAPE-PLD caused a decrease in the total amount of NAPEs by 50-90% with a 1.5-fold increase in the total amount of NAEs, suggesting that the recombinant NAPE-PLD utilizes endogenous NAPE as a substrate in the cell. Since the compositions of NAEs and NAPEs of NAPE-PLD-overexpressing cells and mock-transfected cells were very similar, the enzyme did not appear to discriminate among the N-acyl groups of endogenous NAPEs. These results confirm that overexpressed NAPE-PLD is capable of forming NAEs, including anandamide, in living cells. |
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Authors:
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Yasuo Okamoto; Jun Morishita; Jun Wang; Patricia C Schmid; Randy J Krebsbach; Harald H O Schmid; Natsuo Ueda |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: The Biochemical journal Volume: 389 ISSN: 1470-8728 ISO Abbreviation: Biochem. J. Publication Date: 2005 Jul |
Date Detail:
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Created Date: 2005-06-15 Completed Date: 2005-12-12 Revised Date: 2009-11-18 |
Medline Journal Info:
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Nlm Unique ID: 2984726R Medline TA: Biochem J Country: England |
Other Details:
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Languages: eng Pagination: 241-7 Citation Subset: IM |
Affiliation:
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Department of Biochemistry, Kagawa University School of Medicine, 1750-1 Ikenobe, Miki, Kagawa 761-0793, Japan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals CHO Cells COS Cells Cercopithecus aethiops Cricetinae Gene Expression / genetics Humans Mice Phosphatidylethanolamines / metabolism* Phospholipase D / genetics, metabolism* |
| Chemical | |
Reg. No./Substance:
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0/Phosphatidylethanolamines; EC 3.1.4.4/N-acylphosphatidylethanolamine phospholipase D, mouse; EC 3.1.4.4/Phospholipase D |
| Comments/Corrections | |
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