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.