Abstract
Interferon gamma (IFNγ) is a potent inflammatory and immune cytokine. IFNγ signals via the interferon gamma receptor (IFNGR), which is constitutively bound to Janus Kinase (JAK) 1 and JAK2 via its intracellular domain. These two JAK proteins then initiate the inflammatory signaling cascade. The most potent inhibitor of IFNγ signaling is Suppressor of Cytokine Signaling 1 (SOCS1). SOCS1 negatively regulates IFNγ signaling pathway (and other pathways) by directly inhibiting JAKs. Here, we describe a protocol for the recombinant production and purification of the JAK1 kinase domain and its inhibitor SOCS1, for structural and biochemical studies.
Keywords:
Janus Kinase (JAK); Protein expression; Protein purification; Suppressor of cytokine signaling (SOCS).
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Baculoviridae / genetics
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Baculoviridae / metabolism
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Cells, Cultured
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Humans
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Janus Kinase 1 / genetics
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Janus Kinase 1 / isolation & purification*
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Janus Kinase 1 / metabolism*
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Sf9 Cells
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Signal Transduction
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Suppressor of Cytokine Signaling 1 Protein / genetics
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Suppressor of Cytokine Signaling 1 Protein / isolation & purification*
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Suppressor of Cytokine Signaling 1 Protein / metabolism*
Substances
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Recombinant Proteins
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SOCS1 protein, human
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Suppressor of Cytokine Signaling 1 Protein
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JAK1 protein, human
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Janus Kinase 1