Document Detail

Generation of N-acylphosphatidylethanolamine by members of the phospholipase A/acyltransferase (PLA/AT) family.
MedLine Citation:
PMID:  22825852     Owner:  NLM     Status:  MEDLINE    
Bioactive N-acylethanolamines (NAEs), including N-palmitoylethanolamine, N-oleoylethanolamine, and N-arachidonoylethanolamine (anandamide), are formed from membrane glycerophospholipids in animal tissues. The pathway is initiated by N-acylation of phosphatidylethanolamine to form N-acylphosphatidylethanolamine (NAPE). Despite the physiological importance of this reaction, the enzyme responsible, N-acyltransferase, remains molecularly uncharacterized. We recently demonstrated that all five members of the HRAS-like suppressor tumor family are phospholipid-metabolizing enzymes with N-acyltransferase activity and are renamed HRASLS1-5 as phospholipase A/acyltransferase (PLA/AT)-1-5. However, it was poorly understood whether these proteins were involved in the formation of NAPE in living cells. In the present studies, we first show that COS-7 cells transiently expressing recombinant PLA/AT-1, -2, -4, or -5, and HEK293 cells stably expressing PLA/AT-2 generated significant amounts of [(14)C]NAPE and [(14)C]NAE when cells were metabolically labeled with [(14)C]ethanolamine. Second, as analyzed by liquid chromatography-tandem mass spectrometry, the stable expression of PLA/AT-2 in cells remarkably increased endogenous levels of NAPEs and NAEs with various N-acyl species. Third, when NAPE-hydrolyzing phospholipase D was additionally expressed in PLA/AT-2-expressing cells, accumulating NAPE was efficiently converted to NAE. We also found that PLA/AT-2 was partly responsible for NAPE formation in HeLa cells that endogenously express PLA/AT-2. These results suggest that PLA/AT family proteins may produce NAPEs serving as precursors of bioactive NAEs in vivo.
Toru Uyama; Natsuki Ikematsu; Manami Inoue; Naoki Shinohara; Xing-Hua Jin; Kazuhito Tsuboi; Takeharu Tonai; Akira Tokumura; Natsuo Ueda
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2012-07-23
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  287     ISSN:  1083-351X     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2012 Sep 
Date Detail:
Created Date:  2012-09-17     Completed Date:  2012-12-04     Revised Date:  2013-09-17    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  31905-19     Citation Subset:  IM    
Department of Biochemistry, Kagawa University School of Medicine, 1750-1 Ikenobe, Miki, Kagawa 761-0793, Japan.
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MeSH Terms
Acyltransferases / chemistry*
Base Sequence
COS Cells
Cercopithecus aethiops
Gene Expression Regulation*
HEK293 Cells
HeLa Cells
Molecular Sequence Data
Multigene Family
Peroxisomes / metabolism
Phosphatidylethanolamines / chemistry*
Phospholipase D / metabolism
Phospholipases A / chemistry*
Phospholipids / chemistry
RNA Interference
Tandem Mass Spectrometry / methods
Reg. No./Substance:
0/Phosphatidylethanolamines; 0/Phospholipids; EC 2.3.-/Acyltransferases; EC 3.1.1.-/Phospholipases A; EC D

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