| RRC ID |
56116
|
| 著者 |
Hussain Z, Uyama T, Kawai K, Binte Mustafiz SS, Tsuboi K, Araki N, Ueda N.
|
| タイトル |
Phosphatidylserine-stimulated production of N-acyl-phosphatidylethanolamines by Ca2+-dependent N-acyltransferase.
|
| ジャーナル |
Biochim Biophys Acta Mol Cell Biol Lipids
|
| Abstract |
N-acyl-phosphatidylethanolamine (NAPE) is known to be a precursor for various bioactive N-acylethanolamines including the endocannabinoid anandamide. NAPE is produced in mammals through the transfer of an acyl chain from certain glycerophospholipids to phosphatidylethanolamine (PE) by Ca2+-dependent or -independent N-acyltransferases. The ε isoform of mouse cytosolic phospholipase A2 (cPLA2ε) was recently identified as a Ca2+-dependent N-acyltransferase (Ca-NAT). In the present study, we first showed that two isoforms of human cPLA2ε function as Ca-NAT. We next purified both mouse recombinant cPLA2ε and its two human orthologues to examine their catalytic properties. The enzyme absolutely required Ca2+ for its activity and the activity was enhanced by phosphatidylserine (PS). PS enhanced the activity 25-fold in the presence of 1 mM CaCl2 and lowered the EC50 value of Ca2+ >8-fold. Using a PS probe, we showed that cPLA2ε largely co-localizes with PS in plasma membrane and organelles involved in the endocytic pathway, further supporting the interaction of cPLA2ε with PS in living cells. Finally, we found that the Ca2+-ionophore ionomycin increased [14C]NAPE levels >10-fold in [14C]ethanolamine-labeled cPLA2ε-expressing cells while phospholipase A/acyltransferase-1, acting as a Ca2+-independent N-acyltransferase, was insensitive to ionomycin for full activity. In conclusion, PS potently stimulated the Ca2+-dependent activity and human cPLA2ε isoforms also functioned as Ca-NAT.
|
| 巻・号 |
1863(5)
|
| ページ |
493-502
|
| 公開日 |
2018-5-1
|
| DOI |
10.1016/j.bbalip.2018.02.002
|
| PII |
S1388-1981(18)30022-2
|
| PMID |
29447909
|
| MeSH |
Acyltransferases / chemistry
Acyltransferases / metabolism*
Amino Acid Sequence
Animals
Biosynthetic Pathways / drug effects
COS Cells
Calcium / pharmacology*
Cations, Divalent / pharmacology
Cell Survival / drug effects
Chlorocebus aethiops
Ethanolamines / metabolism
Humans
Ionomycin / pharmacology
Mice
Phosphatidylethanolamines / metabolism*
Phosphatidylserines / metabolism*
Phospholipases A2, Cytosolic / chemistry
Phospholipases A2, Cytosolic / metabolism
Plasmalogens / metabolism
RAW 264.7 Cells
Sequence Homology, Amino Acid
|
| IF |
4.402
|
| 引用数 |
5
|
| リソース情報 |
| ヒト・動物細胞 |
RAW 264(RCB0535) |
