@article{2e880760479d497abf96615af3549d5c,
title = "Lysophospholipases cooperate to mediate lipid homeostasis and lysophospholipid signaling",
abstract = "Abstract Lysophospholipids (LysoPLs) are bioactive lipid species involved in cellular signaling processes and the regulation of cell membrane structure. LysoPLs are metabolized through the action of lysophospholipases, including lysophospholipase A1 (LYPLA1) and lysophospholipase A2 (LYPLA2). A new X-ray crystal structure of LYPLA2 compared with a previously published structure of LYPLA1 demonstrated near-identical folding of the two enzymes; however, LYPLA1 and LYPLA2 have displayed distinct substrate specificities in recombinant enzyme assays. To determine how these in vitro substrate preferences translate into a relevant cellular setting and better understand the enzymes{\textquoteright} role in LysoPL metabolism, CRISPR-Cas9 technology was utilized to generate stable KOs of Lypla1 and/or Lypla2 in Neuro2a cells. Using these cellular models in combination with a targeted lipidomics approach, LysoPL levels were quantified and compared between cell lines to determine the effect of losing lysophospholipase activity on lipid metabolism. This work suggests that LYPLA1 and LYPLA2 are each able to account for the loss of the other to maintain lipid homeostasis in cells; however, when both are deleted, LysoPL levels are dramatically increased, causing phenotypic and morphological changes to the cells.—Wepy, J. A., James J. Galligan, P. J. Kingsley, S. Xu, M. C. Goodman, K. A. Tallman, C. A. Rouzer, and L. J. Marnett. Lysophospholipases cooperate to mediate lipid homeostasis and lysophospholipid signaling.",
keywords = "Brain lipids, Eicosanoids, Fatty acid, Fluorescence microscopy, Lipidomics, Mass spectrometry, Neurons, Prostaglandins, Protein kinases/MAP kinase, X-ray crystallography",
author = "Wepy, {James A.} and Galligan, {James J.} and Kingsley, {Philip J.} and Shu Xu and Goodman, {Michael C.} and Tallman, {Keri A.} and Rouzer, {Carol A.} and Marnett, {Lawrence J.}",
note = "Funding Information: Flow Cytometry experiments were performed in the VMC Flow Cytometry Shared Resource. The VMC Flow Cytometry Shared Resource is supported by the Vanderbilt Ingram Cancer Center (P30 CA68485) and the Vanderbilt Digestive Disease Research Center (DK058404). Antibodies were generated and purified by the Vanderbilt Antibody and Protein Resource. The Vanderbilt Antibody and Protein Resource is supported by the Vanderbilt Institute of Chemical Biology and the Vanderbilt Ingram Cancer Center (P30 CA68485). Imaging experiments were performed in the Vanderbilt High-Throughput Screening Core Facility with assistance provided by Debbie Mi and Joshua Bauer. The HTS core receives support from the Vanderbilt Institute of Chemical Biology and the Vanderbilt-Ingram Cancer Center (P30 CA68485). Statistical analyses were performed with guidance by Yu Shyr, Ph.D. from the Department of Biostatistics at Vanderbilt University. Funding Information: This work was supported by National Institutes of Health Grants CA089450 and T32-ES007028; the Hancock Jr. Memorial Cancer Center Fund; and National Institute of General Medical Sciences Fellowship Ruth L. Kirschstein National Research Service Award 5F31GM120879-02. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the National Institutes of Health. Manuscript received 19 June 2018 and in revised form 5 November 2018. Published, JLR Papers in Press, November 27, 2018 DOI https://doi.org/10.1194/jlr.M087890 Publisher Copyright: Copyright {\textcopyright} 2019 Wepy et al. Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc.",
year = "2019",
doi = "10.1194/jlr.M087890",
language = "English (US)",
volume = "60",
pages = "360--374",
journal = "Journal of Lipid Research",
issn = "0022-2275",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "2",
}