Fatty Acyl Availability Modulates Cardiolipin Composition and Alters Mitochondrial Function in HeLa Cells

LC-MS/MS raw data and quantified cardiolipin data for the manuscript 'Fatty Acyl Availability Modulates Cardiolipin Composition and Alters Mitochondrial Function in HeLa Cells '.

Cardiolipin analysis was performed as described in Oemer et al, 2018, PNAS. Briefly, sample material was homogenized in PBS and lipids were extracted following the Folch method with internal standard (CL(14:0)4, 0.5 µM). Lipid extracts were dissolved in HPLC starting condition and subjected to HPLC-MS/MS analysis. Separation was achieved by reversed-phase HPLC with an Agilent Poroshell 120 EC-C8 2.7 µm 2.1x100mm column (Agilent Technologies, Santa Clara, USA) on a Dionex Ultimate 3000 HPLC (Thermo Fisher Scientific Inc, Waltham, USA, 50°C column oven, 0.4 µl/min flow) with running solvent A (60/40 Acetonitrile/H2O, 10 mM ammonium formate, 0.2% formic acid) and running solvent B (90/10 Isopropanol/Acetonitrile, 10 mM ammonium formate, 0.2% formic acid). Analytes were measured using a LTQ Velos MS (Thermo Fisher Scientific Inc, Waltham, USA) operated in negative ESI mode (3.8kV, 275°C capillary temperature, 460 - 1650 m/z) and data-dependent MS2 acquisition. Thermo raw data was converted to open-source MZML format and Peaks were integrated in MZmine2. Identification was based on a combination of accurate mass, (relative) retention times, and fragmentation spectra, compared to a library of standards. Normalization, quantification and data analysis was performed by an in-house pipeline in R.

本数据集对应论文《脂肪酸酰基可用性调控HeLa细胞中心磷脂组成并改变线粒体功能（Fatty Acyl Availability Modulates Cardiolipin Composition and Alters Mitochondrial Function in HeLa Cells）》，包含其液相色谱-串联质谱（Liquid Chromatography-Tandem Mass Spectrometry, LC-MS/MS）原始数据与定量心磷脂（cardiolipin）数据。心磷脂分析参照Oemer等人2018年发表于《美国国家科学院院刊（Proceedings of the National Academy of Sciences, PNAS）》的方法进行。简言之，将样品材料在磷酸盐缓冲液（PBS）中匀浆，随后采用Folch法结合内标（CL(14:0)4，0.5 微摩尔）提取脂质。将脂质提取物溶于高效液相色谱（High Performance Liquid Chromatography, HPLC）初始流动相后，进行HPLC-MS/MS分析。色谱分离采用反相高效液相色谱体系，搭载Dionex Ultimate 3000 HPLC系统（赛默飞世尔科技公司，美国沃尔瑟姆），使用Agilent Poroshell 120 EC-C8 2.7 µm 2.1×100 mm色谱柱（安捷伦科技公司，美国圣克拉拉）；柱温箱温度设为50°C，流速为0.4 微升/分钟，流动相A为60/40乙腈/水溶液（含10 毫摩尔甲酸铵、0.2%甲酸），流动相B为90/10异丙醇/乙腈溶液（含10 毫摩尔甲酸铵、0.2%甲酸）。待测物采用LTQ Velos质谱仪（赛默飞世尔科技公司，美国沃尔瑟姆）以负电喷雾电离（ESI）模式检测，参数设置为：喷雾电压3.8 kV，毛细管温度275°C，扫描范围460–1650 质荷比，并采用数据依赖型二级质谱（MS2）采集模式。将Thermo原始数据转换为开源格式MZML，通过MZmine2软件完成峰积分。心磷脂鉴定结合精确质量、（相对）保留时间与碎裂光谱，并与标准品谱库比对完成。标准化处理、定量分析与数据统计均通过R语言自研流程实现。