title: Phosphatidic acid-mediated activation and translocation to the cell surface of sialidase NEU3, promoting signaling for cell migration.
abstract: The plasma membrane-associated sialidase NEU3 plays crucial roles in regulation of transmembrane signaling, and its aberrant up-regulation in various cancers contributes to malignancy. However, it remains uncertain how NEU3 is naturally activated and locates to plasma membranes, because of its Triton X-100 requirement for the sialidase activity in vitro and its often changing subcellular location. Among phospholipids examined, we demonstrate that phosphatidic acid (PA) elevates its sialidase activity 4 to 5 times at 50 μM in vitro at neutral pH and promotes translocation to the cell surface and cell migration through Ras-signaling in HeLa and COS-1 cells. NEU3 was found to interact selectively with PA as assessed by phospholipid array, liposome coprecipitation, and ELISA assays and to colocalize with phospholipase D (PLD) 1 in response to epidermal growth factor (EGF) or serum stimulation. Studies using tagged NEU3 fragments with point mutations identified PA- and calmodulin (CaM)-binding sites around the N terminus and confirmed its participation in translocation and catalytic activity. EGF induced PLD1 activation concomitantly with enhanced NEU3 translocation to the cell surface, as assessed by confocal microscopy. These results suggest that interactions of NEU3 with PA produced by PLD1 are important for regulation of transmembrane signaling, this aberrant acceleration probably promoting malignancy in cancers.
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Kazuhiro
authors: K
authors: Shiozaki
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Kohta
authors: K
authors: Takahashi
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Masahiro
authors: M
authors: Hosono
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Kazunori
authors: K
authors: Yamaguchi
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Keiko
authors: K
authors: Hata
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Momo
authors: M
authors: Shiozaki
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Rosaria
authors: R
authors: Bassi
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Alessandro
authors: A
authors: Prinetti
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Sandro
authors: S
authors: Sonnino
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy.
authors: Kazuo
authors: K
authors: Nitta
authors: *Faculty of Fisheries and The United Graduate School of Agricultural Science, Kagoshima University, Kagoshima, Kagoshima, Japan; Division of Cancer Glycosylation Research and Division of Cell Recognition Study, Tohoku Pharmaceutical University, Sendai, Miyagi, Japan; Miyagi Cancer Center Research Institute, Natori, Miyagi, Japan; and Department of Medical Biotechnology and Translational Medicine, University of Milan, Segrate, Milan, Italy tmiyagi@tohoku-pharm.ac.jp.
authors: Taeko
authors: T
authors: Miyagi
mesh_terms: D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism*; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004789:Enzyme Activation; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D009439:Neuraminidase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D010712:Phosphatidic Acids / Q000494:pharmacology*; D010739:Phospholipase D / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport / Q000187:drug effects*; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D010712:Phosphatidic Acids; D034741:RNA, Small Interfering; D010739:Phospholipase D; C106801:phospholipase D1; C465436:Neu3 protein, human; D009439:Neuraminidase
keywords: PLD1; cancer; gangliosides; phospholipids; sialic acid