sn-Position-Resolved Quantification of Aminophospholipids by Isotopic N,N-Dimethyl Leucine Labeling and High-Resolution Ion Mobility Mass Spectrometry

IF 6.7 1区 化学 Q1 CHEMISTRY, ANALYTICAL
Shuling Xu, Zhijun Zhu, Ting-Jia Gu, Zicong Wang, Daniel G. Delafield, Michael J. Rigby, Gaoyuan Lu, Min Ma, Peng-Kai Liu, Luigi Puglielli, Lingjun Li
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引用次数: 0

Abstract

Aminophospholipids (APLs), composed of phosphatidylethanolamines (PEs) and phosphatidylserines (PSs), are vital components of mammalian cell membranes and lipoproteins, participating in both homeostasis and cellular signaling. Their structural changes, including the permutation of fatty acid connectivity (sn-positions), due to dysfunctional metabolic processes have been linked to many diseases. However, the accurate quantification of APLs with unambiguous fatty acyl assignment through routine label-free LC–MS/MS lipidomic analysis remains a major challenge. In this study, we explore the functionalization of the free primary amine groups of APLs using amine-reactive isotopic N,N-dimethyl leucine (iDiLeu) and employ high-resolution ion mobility MS (IM-MS) to develop a novel method for sensitive discernment and accurate quantification of APL sn-isomers. With high-resolution demultiplexing (HRdm) providing IM resolving power >200, labeled sn-isomeric pairs of APLs (ΔCCS ≈ 1%) demonstrate excellent, near baseline separation. In addition to greatly enhanced sensitivity, 5-plex iDiLeu labeling enables the construction of an internal 4-point calibration curve and therefore absolute quantification of APL sn-isomers in a single run. This strategy enabled precise annotation and quantification of 239 APLs including 60 pairs of sn-isomers in the mouse cortex. Additionally, we were able to find ratio changes in multiple APL sn-isomer pairs between wild type and APP/PS1 Alzheimer’s disease (AD) model mice at different ages, indicating their strong correlation to AD progression. This strategy could provide universal utility in unraveling the alteration of APL sn-isomers, which have long been considered as the “dark matter” of traditional lipidomic analyses, leading to more precise elucidation of molecular mechanisms of various diseases.

Abstract Image

用同位素N、N-二甲基亮氨酸标记和高分辨率离子迁移质谱法进行N-位置分辨定量分析氨基磷脂
由磷脂酰乙醇胺(PEs)和磷脂酰丝氨酸(ps)组成的氨基磷脂(api)是哺乳动物细胞膜和脂蛋白的重要组成部分,参与体内平衡和细胞信号传导。它们的结构变化,包括脂肪酸连接(sn位置)的排列,由于功能失调的代谢过程与许多疾病有关。然而,通过常规无标签LC-MS /MS脂质组学分析准确定量具有明确脂肪酰基分配的api仍然是一个主要挑战。在这项研究中,我们利用胺反应同位素N,N-二甲基亮氨酸(iDiLeu)探索了APL中游离氨基的功能化,并采用高分辨率离子迁移率质谱(IM-MS)建立了一种灵敏识别和准确定量APL sn异构体的新方法。高分辨率解复用(HRdm)提供了IM分辨率>;200,标记的n-异构体对的api (ΔCCS≈1%)表现出优异的,接近基线的分离。除了大大提高灵敏度外,5-plex iDiLeu标记可以构建内部4点校准曲线,因此可以在一次运行中对APL sn-异构体进行绝对定量。该策略能够精确地注释和定量小鼠皮质中的239个api,包括60对sn-异构体。此外,我们能够发现野生型和APP/PS1 AD模型小鼠在不同年龄的多个APL sn-异构体对的比例变化,表明它们与AD的进展有很强的相关性。这种策略可以为揭示APL sn-异构体的改变提供普遍的实用价值,这一直被认为是传统脂质组学分析的“暗物质”,从而更精确地阐明各种疾病的分子机制。
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来源期刊
Analytical Chemistry
Analytical Chemistry 化学-分析化学
CiteScore
12.10
自引率
12.20%
发文量
1949
审稿时长
1.4 months
期刊介绍: Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.
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