title: Src Tyrosine Kinase Activation by 4-Hydroxynonenal Upregulates p38, ERK/AP-1 Signaling and COX-2 Expression in YPEN-1 Cells.
abstract: 4-Hydroxynonenal (4-HNE), a major end product of lipid peroxidation, is highly reactive and involved in various cellular processes, such as inflammatory signaling. However, to date, the mechanistic roles of 4-HNE in inflammatory signaling related to protein tyrosine kinases have not been elucidated. In the present study, we investigated the interaction between 4-HNE and Src (a non-receptor tyrosine kinase) for its involvement in the molecular modulation of the inflammatory signaling pathway utilizing the YPEN-1 cell system. Immunoprecipitation experiments showed that 4-HNE phosphorylates (activates) Src at Tyr416 via adduct formation. In addition, LC-MS/MS and a docking simulation model revealed an addiction site at the Cys248 residue of Src, resulting in the stimulation of downstream p38, ERK/AP-1 and cyclooxygenase-2 (COX-2) signaling in YPEN-1 cells. The role of 4-HNE-activated Src in downstream inflammatory signaling was further investigated using dasatinib (a Src inhibitor) and by siRNA knockdown of Src. p38 and ERK were directly regulated by Src, as revealed by immunoblotting of the phosphorylated forms of mitogen-activated protein kinases (MAPKs), which are key elements in the signaling transduction pathway initiated by Src. The study also shows that Src modulates the HNE-enhanced activation of AP-1 and the expression of COX-2 (a target gene of AP-1). Together, the results of this study show that 4-HNE stimulates Src tyrosine kinase in activation of the inflammation process.
authors.affiliation: Molecular Inflammation Research Center for Aging Intervention (MRCA), Department of Pharmacy, Pusan National University, Busan, Republic of Korea.
authors.forename: Eun Ji
authors.initials: EJ
authors.lastname: Jang
authors.affiliation: Molecular Inflammation Research Center for Aging Intervention (MRCA), Department of Pharmacy, Pusan National University, Busan, Republic of Korea; Interdisciplinary Research Program of Bioinformatics and Longevity Science, Pusan National University, Busan 609-735, Republic of Korea.
authors.forename: Hyoung Oh
authors.initials: HO
authors.lastname: Jeong
authors.affiliation: Molecular Inflammation Research Center for Aging Intervention (MRCA), Department of Pharmacy, Pusan National University, Busan, Republic of Korea; Interdisciplinary Research Program of Bioinformatics and Longevity Science, Pusan National University, Busan 609-735, Republic of Korea.
authors.forename: Daeui
authors.initials: D
authors.lastname: Park
authors.affiliation: Molecular Inflammation Research Center for Aging Intervention (MRCA), Department of Pharmacy, Pusan National University, Busan, Republic of Korea.
authors.forename: Dae Hyun
authors.initials: DH
authors.lastname: Kim
authors.affiliation: Molecular Inflammation Research Center for Aging Intervention (MRCA), Department of Pharmacy, Pusan National University, Busan, Republic of Korea.
authors.forename: Yeon Ja
authors.initials: YJ
authors.lastname: Choi
authors.affiliation: Molecular Inflammation Research Center for Aging Intervention (MRCA), Department of Pharmacy, Pusan National University, Busan, Republic of Korea.
authors.forename: Ki Wung
authors.initials: KW
authors.lastname: Chung
authors.affiliation: Molecular Inflammation Research Center for Aging Intervention (MRCA), Department of Pharmacy, Pusan National University, Busan, Republic of Korea.
authors.forename: Min Hi
authors.initials: MH
authors.lastname: Park
authors.affiliation: Department of Physiology, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900, United States of America.
authors.forename: Byung Pal
authors.initials: BP
authors.lastname: Yu
authors.affiliation: Molecular Inflammation Research Center for Aging Intervention (MRCA), Department of Pharmacy, Pusan National University, Busan, Republic of Korea.
authors.forename: Hae Young
authors.initials: HY
authors.lastname: Chung
mesh_terms: D000447:Aldehydes / Q000737:chemistry*; D000818:Animals; D001665:Binding Sites; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000378:metabolism; D002853:Chromatography, Liquid; D051546:Cyclooxygenase 2 / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D007249:Inflammation / Q000378:metabolism; D062105:Molecular Docking Simulation; D018384:Oxidative Stress; D010766:Phosphorylation; D034741:RNA, Small Interfering / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry; D018808:Transcription Factor AP-1 / Q000378:metabolism*; D015854:Up-Regulation; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: D000447:Aldehydes; D034741:RNA, Small Interfering; D018808:Transcription Factor AP-1; D051546:Cyclooxygenase 2; C489691:Ptgs2 protein, rat; D019061:src-Family Kinases; D048049:Extracellular Signal-Regulated MAP Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C027576:4-hydroxy-2-nonenal