title: Quantitative structural characterization of phosphatidylinositol phosphates from biological samples.
abstract: The aspects of cellular metabolism controlled by phosphatidylinositol phosphates (PtdInsPs) have been broadly expanded, and these phospholipids have drawn tremendous attention as pleiotropic signaling molecules. PtdInsPs analysis using LC/MS/MS has remained challenging due to the strong hydrophilicity of these lipids. Multiple reaction monitoring (MRM) or a neutral loss scan has been performed to quantitatively measure PtdInsPs after chemical derivatization on the phosphate groups of inositol moieties. Only predefined PtdInsPs can be measured in MRM mode, and fatty acyl compositions of sn-1 and sn-2 positions of PtdInsPs cannot be obtained from a neutral loss scan. In our present study, we developed a simple LC/MS/MS method for structural identification of sn-1 and sn-2 fatty acids of PtdInsPs and their relative quantitation. Precursor ion scans of sn-1 monoacylglycerols (MAGs) of PtdInsPs provided structural information about the lipids, and ammonium adduction enhanced signal intensities of PtdInsPs. The relative amount of observed PtdInsPs in biological samples could be compared using chromatographic peak areas from the neutral loss scans. Using precursor ion scans of sn-1 MAG and neutral loss scans of headgroups, major PtdInsPs in cells and tissues were successfully identified with structural information of sn-1 and sn-2 fatty acids, and their relative amounts in different samples were compared.
authors.affiliation: Biomedical Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea.
authors.forename: Su Hee
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Biomedical Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea.
authors.forename: Ha Eun
authors.initials: HE
authors.lastname: Song
authors.affiliation: Biomedical Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea.
authors.forename: Su Jung
authors.initials: SJ
authors.lastname: Kim
authors.affiliation: Biomedical Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea.|Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea.
authors.forename: Dong Cheol
authors.initials: DC
authors.lastname: Woo
authors.affiliation: Division of Biomedical Sciences, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea.
authors.forename: Suhwan
authors.initials: S
authors.lastname: Chang
authors.affiliation: School of Biological Sciences, University of Ulsan, Ulsan 44610, Republic of Korea.
authors.forename: Woo Gyun
authors.initials: WG
authors.lastname: Choi
authors.affiliation: School of Biological Sciences, University of Ulsan, Ulsan 44610, Republic of Korea.
authors.forename: Mi Jeong
authors.initials: MJ
authors.lastname: Kim
authors.affiliation: School of Biological Sciences, University of Ulsan, Ulsan 44610, Republic of Korea.
authors.forename: Sung Hoon
authors.initials: SH
authors.lastname: Back
authors.affiliation: Biomedical Research Center, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea yoohyunju@amc.seoul.kr.|Department of Convergence Medicine, Asan Institute for Life Sciences, Asan Medical Center, University of Ulsan College of Medicine, Seoul 05505, Republic of Korea.
authors.forename: Hyun Ju
authors.initials: HJ
authors.lastname: Yoo
mesh_terms: D002853:Chromatography, Liquid / Q000379:methods; D005227:Fatty Acids / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D018129:Phosphatidylinositol Phosphates / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D005227:Fatty Acids; D018129:Phosphatidylinositol Phosphates; D010743:Phospholipids
keywords: fatty acids; high-performance liquid chromatography; lipidomics; phosphoinositides; tandem mass spectrometry