Hänninen Satu, Somerharju Pentti, Hermansson Martin
Department of Biochemistry and Developmental Biology, Medical Faculty, University of Helsinki, Helsinki, Finland.
Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark.
Bio Protoc. 2017 May 5;7(9):e2268. doi: 10.21769/BioProtoc.2268.
Glycerophospholipids consist of a glycerophosphate backbone to which are esterified two acyl chains and a polar head group. The head group ., choline, ethanolamine, serine or inositol) defines the glycerophospholipid class, while the acyl chains together with the head group define the glycerophospholipid molecular species. Stable heavy isotope (., deuterium)-labeled head group precursors added to the culture medium incorporate efficiently into glycerophospholipids of mammalian cells, which allows one to determine the rates of synthesis, acyl chain remodeling or turnover of the individual glycerophospholipids using mass spectrometry. This protocol describes how to study the metabolism of the major mammalian glycerophospholipids ., phosphatidylcholines, phosphatidylethanolamines, phosphatidylserines and phosphatidylinositols with this approach.
甘油磷脂由一个甘油磷酸主链组成，两条酰基链和一个极性头部基团通过酯化作用连接在该主链上。头部基团（如胆碱、乙醇胺、丝氨酸或肌醇）决定了甘油磷脂的类别，而酰基链与头部基团共同决定了甘油磷脂的分子种类。添加到培养基中的稳定重同位素（如氘）标记的头部基团前体能够有效地掺入哺乳动物细胞的甘油磷脂中，这使得人们可以使用质谱法来确定各个甘油磷脂的合成速率、酰基链重塑或周转情况。本方案描述了如何用这种方法研究主要的哺乳动物甘油磷脂（如磷脂酰胆碱、磷脂酰乙醇胺、磷脂酰丝氨酸和磷脂酰肌醇）的代谢。
