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: Phattara-orn Havanapan, Rattiyaporn Kanlaya, Apichai Bourchookarn, Chartchai Krittanai, Visith Thongboonkerd
• Journal: Journal of proteome research (J Proteome Res) Vol. 8 Issue 5 Pg. 2476-83 (May 2009) ISSN: 1535-3893 [Print] United States
• PMID: 19284748 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Serine
• Hemocyanins
• Extracellular Signal-Regulated MAP Kinases
• Mitogen-Activated Protein Kinase 1
• Mitogen-Activated Protein Kinase 3

Topics

• Amino Acid Sequence
• Animals
• Binding Sites
• Blotting, Western
• Electrophoresis, Gel, Two-Dimensional
• Extracellular Signal-Regulated MAP Kinases (metabolism)
• Hemocyanins (analysis, chemistry, metabolism)
• Hemocytes (metabolism)
• Host-Pathogen Interactions
• Immunity, Innate
• Immunoprecipitation
• Mitogen-Activated Protein Kinase 1 (metabolism)
• Mitogen-Activated Protein Kinase 3 (metabolism)
• Molecular Sequence Data
• Penaeidae (immunology, metabolism, virology)
• Phosphorylation
• Protein Binding
• RNA Viruses (immunology, physiology)
• Serine (metabolism)
• Spectrometry, Mass, Electrospray Ionization