title: Gintonin, newly identified compounds from ginseng, is novel lysophosphatidic acids-protein complexes and activates G protein-coupled lysophosphatidic acid receptors with high affinity.
abstract: Recently, we isolated a subset of glycolipoproteins from Panax ginseng, that we designated gintonin, and demonstrated that it induced [Ca2+]i transients in cells via G protein-coupled receptor (GPCR) signaling pathway(s). However, active components responsible for Ca2+ mobilization and the corresponding receptor(s) were unknown. Active component(s) for [Ca2+]i transients of gintonin were analyzed by liquid chromatography-electrospray ionization-tandem mass spectrometry and ion-mobility mass spectrometry, respectively. The corresponding receptor(s)were investigated through gene expression assays. We found that gintonin contains LPA C18:2 and other LPAs. Proteomic analysis showed that ginseng major latex-like protein and ribonuclease-like storage proteins are protein components of gintonin. Gintonin induced [Ca2+]i transients in B103 rat neuroblastoma cells transfected with human LPA receptors with high affinity in order of LPA2 >LPA5 > LPA1 > LPA3 > LPA4. The LPA1/LPA3 receptor antagonist Ki16425 blocked gintonin action in cells expressing LPA1 or LPA3. Mutations of binding sites in the LPA3 receptor attenuated gintonin action. Gintonin acted via pertussis toxin (PTX)-sensitive and -insensitive G protein-phospholipase C (PLC)-inositol 1,4,5-trisphosphate (IP3)-Ca2+ pathways. However, gintonin had no effects on other receptors examined. In human umbilical vein endothelial cells (HUVECs) gintonin stimulated cell proliferation and migration. Gintonin stimulated ERK1/2 phosphorylation. PTX blocked gintonin-mediated migration and ERK1/2 phosphorylation. In PC12 cells gintonin induced morphological changes, which were blocked by Rho kinase inhibitorY-27632. Gintonin contains GPCR ligand LPAs in complexes with ginseng proteins and could be useful in the development of drugs targeting LPA receptors.
authors.affiliation: Ginsentology Research Laboratory and Department of Physiology, and Bio/Molecular Informatics Center, College of Veterinary Medicine, Konkuk University, Seoul 143-701, Korea.
authors.forename: Sung Hee
authors.initials: SH
authors.lastname: Hwang
authors.forename: Tae-Joon
authors.initials: TJ
authors.lastname: Shin
authors.forename: Sun-Hye
authors.initials: SH
authors.lastname: Choi
authors.forename: Hee-Jung
authors.initials: HJ
authors.lastname: Cho
authors.forename: Byung-Hwan
authors.initials: BH
authors.lastname: Lee
authors.forename: Mi Kyung
authors.initials: MK
authors.lastname: Pyo
authors.forename: Jun-Ho
authors.initials: JH
authors.lastname: Lee
authors.forename: Jiyeon
authors.initials: J
authors.lastname: Kang
authors.forename: Hyeon-Joong
authors.initials: HJ
authors.lastname: Kim
authors.forename: Chan-Woo
authors.initials: CW
authors.lastname: Park
authors.forename: Ho-Chul
authors.initials: HC
authors.lastname: Shin
authors.forename: Seung-Yeol
authors.initials: SY
authors.lastname: Nah
mesh_terms: D000818:Animals; D020013:Calcium Signaling / Q000235:genetics; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D019204:GTP-Binding Proteins / Q000378:metabolism*; D020869:Gene Expression Profiling; D006023:Glycoproteins / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D007555:Isoxazoles / Q000494:pharmacology; D008246:Lysophospholipids / Q000737:chemistry; D058990:Molecular Targeted Therapy; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D016716:PC12 Cells; D005894:Panax; D037342:Pertussis Toxin / Q000494:pharmacology; D010936:Plant Extracts / Q000737:chemistry*; D010940:Plant Proteins / Q000737:chemistry; D011422:Propionates / Q000494:pharmacology; D011485:Protein Binding / Q000235:genetics; D051381:Rats; D049368:Receptors, Lysophosphatidic Acid / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D019076:Transgenes / Q000235:genetics
chemical_list: C477898:3-(4-(4-((1-(2-chlorophenyl)ethoxy)carbonyl amino)-3-methyl-5-isoxazolyl) benzylsulfanyl) propanoic acid; D006023:Glycoproteins; D007555:Isoxazoles; D008246:Lysophospholipids; D010936:Plant Extracts; D010940:Plant Proteins; D011422:Propionates; D049368:Receptors, Lysophosphatidic Acid; C000603040:gintonin; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins