Recent Advances on Mass Spectrometry Analysis of Nitrated Phospholipids
Recent Advances on Mass Spectrometry Analysis of Nitrated Phospholipids
复制标题
DOI：
10.1021/acs.analchem.5b03407
复制
发表时间：
2016-03-01
影响因子：
7.4
通讯作者：
Domingues, M. Rosario M.
中科院分区：
文献类型：
--
作者：
Melo, Tania;Domingues, Pedro;Domingues, M. Rosario M.
近年来，硝基脂肪酸(NO2-FA)作为一种在氮氧化胁迫下形成的信号分子受到越来越多的关注。NO2-FA在体内以游离形式被检测到，尽管假设它们也可能被酯化成磷脂(PL)。然而，到目前为止，关于硝基磷脂(NO2-PL)的性质、来源和作用的讨论还很少。本研究的目的是建立一种基于质谱仪(MS)的方法，该方法可以识别生命系统中发现的三种主要PL类的硝基烯烃衍生物：磷脂酰胆碱(PC)、磷脂酰乙醇胺(PES)和磷脂酰丝氨酸(PSS)。NO2-PLs是由NO2BF4在疏水环境中模拟生物体系产生的。然后用电喷雾电离(ESI-MS)和电喷雾电离-MS联用亲水作用液相色谱(HILIC)检测NO2-PLs。用串联MS进一步分析了正离子模式(对于所有PL类为[M+H](+)离子)和负离子模式(对于PES和PSS为[M-H](-)离子，对于PCS为[M+OAc](-)离子)。所有NO2-PL的典型MS/MS裂解模式包括HNO2的中性损失，极性头基和HNO2共同损失的产物离子，[NO2-FA+H](+)和[NO2-FA-H](-)产物离子，以及硝基附近脂肪酸主链上的裂解，使其定位于FA烷基内。用改进的MS方法从一种典型的1型糖尿病动物模型中鉴定心肌线粒体中的NO2-PL。我们鉴定了9种NO2-PC和1种NO2-PE。这些发现的生理学相关性仍不清楚。
In recent years, there has been an increasing interest in nitro fatty acids (NO2-FA) as signaling molecules formed under nitroxidative stress. NO2-FA were detected in vivo in a free form, although it is assumed that they may also be esterified to phospholipids (PL). Nevertheless, insufficient discussion about the nature, origin, or role of nitro phospholipids (NO2-PL) was reported up to now. The aim of this study was to develop a mass spectrometry (MS) based approach which allows identifying nitroalkenes derivatives of three major PL classes found in living systems: phosphatidylcholines (PCs), phosphatidylethanolamine (PEs), and phosphatidylserines (PSs). NO2-PLs were generated by NO2BF4 in hydrophobic environment, mimicking biological systems. The NO2-PLs were then detected by electrospray ionization (ESI-MS) and ESI-MS coupled to hydrophilic interaction liquid chromatography (HILIC). Identified NO2-PLs were further analyzed by tandem MS in positive (as [M + H](+) ions for all PL classes) and negative-ion mode (as [M - H](-) ions for PEs and PSs and [M + OAc](-) ions for PCs). Typical MS/MS fragmentation pattern of all NO2-PL included a neutral loss of HNO2, product ions arising from the combined loss of polar headgroup and HNO2, [NO2-FA + H](+) and [NO2-FA - H](-) product ions, and cleavages on the fatty acid backbone near the nitro group, allowing its localization within the FA akyl chain. Developed MS method was used to identify NO2-PL in cardiac mitochondria from a well-characterized animal model of type 1 diabetes mellitus. We identified nine NO2-PCs and one NO2-PE species. The physiological relevance of these findings is still unknown.