RRC ID |
65067
|
著者 |
Hama K, Fujiwara Y, Tabata H, Takahashi H, Yokoyama K.
|
タイトル |
Comprehensive Quantitation Using Two Stable Isotopically Labeled Species and Direct Detection of N-Acyl Moiety of Sphingomyelin.
|
ジャーナル |
Lipids
|
Abstract |
Sphingomyelin (ceramide-phosphocholine, CerPCho) is a common sphingolipid in mammalian cells and is composed of phosphorylcholine and ceramide as polar and hydrophobic components, respectively. In this study, a qualitative liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS/MS) analysis is proposed in which CerPCho structures were assigned based on product ion spectra corresponding to sphingosylphosphorylcholine and N-acyl moieties. From MS/MS/MS analysis of CerPCho, we observed product ion spectra of the N-acyl fatty acids as [RCO2]- ions as well as sphingosylphosphorylcholine. A calibration curve for CerPCho was constructed using two stable isotopically labeled CerPCho species and then used to quantify the CerPCho species in HeLa cells as a proof-of-principle study. The present study proposes an accurate method for quantifying and assigning structures to each CerPCho species in crude biologic samples by LC-ESI-MS/MS/MS analysis.
|
巻・号 |
52(9)
|
ページ |
789-799
|
公開日 |
2017-9-1
|
DOI |
10.1007/s11745-017-4279-5
|
PII |
10.1007/s11745-017-4279-5
|
PMID |
28770378
|
PMC |
PMC5583271
|
MeSH |
Acetyltransferases / genetics
Acetyltransferases / metabolism
Acylation
Chromatography, Liquid
Fatty Acid Elongases
HeLa Cells
Humans
Spectrometry, Mass, Electrospray Ionization
Sphingomyelins / analysis*
Sphingomyelins / metabolism
Tandem Mass Spectrometry
Up-Regulation
|
IF |
1.919
|
リソース情報 |
ヒト・動物細胞 |
HeLa(RCB0007) |