title: C-terminal hemocyanin from hemocytes of Penaeus vannamei interacts with ERK1/2 and undergoes serine phosphorylation.
abstract: To understand molecular immune response of Penaeus vannamei during Taura syndrome virus (TSV) infection, expression and functional proteomics studies were performed on hemocyanin, which is a major abundant protein in shrimp hemocytes. Two-dimensional electrophoresis (2-DE) revealed up-regulation of several C-terminal fragments of hemocyanin, whereas the N-terminal fragments were down-regulated during TSV infection. 2-D Western blot analysis showed that the C-terminal hemocyanin fragments had more acidic isoelectric points (pI), whereas the N-terminal fragments had less acidic pI. Further analysis by NetPhos showed a greater number of serine phosphorylation sites in the C-terminal hemocyanin. Additionally, motif scan using Scansite revealed ERK D-domain, which is required for activation of ERK1/2 effector kinase, as a kinase-binding site at the 527th valine in the C-terminal hemocyanin, whereas neither motif nor functional domain was found in the N-terminus. Co-immunoprecipitation confirmed the interaction between the C-terminal hemocyanin and ERK1/2. 1-D Western blot analysis showed that ERK1/2 was also up-regulated during TSV infection. Our findings demonstrate for the first time that ERK1/2 signaling pathway may play an important role in molecular immune response of P. vannamei upon TSV infection through its interaction with the C-terminal hemocyanin.
authors.affiliation: Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Phattara-orn
authors.initials: PO
authors.lastname: Havanapan
authors.forename: Rattiyaporn
authors.initials: R
authors.lastname: Kanlaya
authors.forename: Apichai
authors.initials: A
authors.lastname: Bourchookarn
authors.forename: Chartchai
authors.initials: C
authors.lastname: Krittanai
authors.forename: Visith
authors.initials: V
authors.lastname: Thongboonkerd
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D015180:Electrophoresis, Gel, Two-Dimensional; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism*; D006433:Hemocyanins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D006434:Hemocytes / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D007113:Immunity, Innate; D047468:Immunoprecipitation; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000276:immunology / Q000378:metabolism* / Q000821:virology; D010766:Phosphorylation; D011485:Protein Binding; D012328:RNA Viruses / Q000276:immunology / Q000502:physiology; D012694:Serine / Q000378:metabolism*; D021241:Spectrometry, Mass, Electrospray Ionization
chemical_list: D012694:Serine; D006433:Hemocyanins; D048049:Extracellular Signal-Regulated MAP Kinases; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3