Abstract
Type II IFN exists as a single molecule (IFN-gamma) in contrast to type I IFN, of which there are a number of different forms. IFN-gamma is involved both directly and indirectly in antiviral activity, stimulation of bactericidal activity, antigen presentation and activation of macrophages. Recently IFN-gamma was cloned from a salmonid fish, the rainbow trout and a functional recombinant protein produced exhibited IFN-gamma activity. This recombinant IFN-gamma was used to stimulate an Atlantic salmon cell line, SHK-1, to monitor the changes in protein expression by proteomic analysis 24 h after stimulation compared to unstimulated control cells. An SHK-1 cell proteome map was developed and proteins altered in abundance by the IFN-gamma stimulation were identified. Under the analytical conditions used, 22 proteins were found to be altered in abundance, 15 increased and 7 decreased. Several proteins were excised from the gel and identified, following trypsin digestion and MALDI-MS/MS/LC-MS and database interrogation. Transcriptional analysis of five mRNAs encoding proteins increased in abundance by IFN-gamma in the proteome analysis was determined by real-time PCR. We assessed the correlation between gene expression and change in abundance of proteins for these genes.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / genetics
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Adenosine Triphosphatases / metabolism
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Animals
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Cell Line
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Chemokine CXCL10
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Chemokines, CXC / genetics
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Chemokines, CXC / metabolism
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Down-Regulation / drug effects
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Electrophoresis, Gel, Two-Dimensional
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Endoplasmic Reticulum Chaperone BiP
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Eukaryotic Initiation Factor-2 / genetics
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Eukaryotic Initiation Factor-2 / metabolism
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Fish Proteins / genetics
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Fish Proteins / metabolism*
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GTP-Binding Proteins / genetics
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GTP-Binding Proteins / metabolism
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Gene Expression / drug effects
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Heat-Shock Proteins / genetics
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Heat-Shock Proteins / metabolism
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Interferon Type I / genetics
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Interferon-gamma / pharmacology*
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Lectins, C-Type / genetics
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Lectins, C-Type / metabolism
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Macrophages / cytology
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Macrophages / drug effects
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Macrophages / metabolism
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Mixed Function Oxygenases / genetics
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Mixed Function Oxygenases / metabolism
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Molecular Chaperones / genetics
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Molecular Chaperones / metabolism
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Myxovirus Resistance Proteins
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Proteome / genetics
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Proteome / metabolism*
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Recombinant Proteins
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Salmo salar / genetics
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Salmo salar / metabolism*
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Tandem Mass Spectrometry
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Up-Regulation / drug effects
Substances
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Chemokine CXCL10
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Chemokines, CXC
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Endoplasmic Reticulum Chaperone BiP
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Eukaryotic Initiation Factor-2
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Fish Proteins
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Heat-Shock Proteins
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Interferon Type I
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Lectins, C-Type
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Molecular Chaperones
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Myxovirus Resistance Proteins
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Proteome
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Recombinant Proteins
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Interferon-gamma
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Mixed Function Oxygenases
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Adenosine Triphosphatases
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GTP-Binding Proteins