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Expression and Purification of JAK1 and SOCS1 for Structural and Biochemical Studies

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Book cover Inflammation and Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1725))

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.

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Acknowledgments

This work was supported by the Cancer Council Victoria (Grant-in-aid 1065180) and the National Health and Medical Research Council (NHMRC) Australia (Project grant #112299, Program grant #1113577), an NHMRC IRIISS grant 361646, and a Victorian State Government Operational Infrastructure Scheme grant. J.J.B. is supported by an NHMRC fellowship and NPDL by an Australian Postgraduate Award.

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Authors and Affiliations

  1. The Walter and Eliza Hall Institute of Medical Research, Parkville, Melbourne, VIC, Australia

    Nicholas P. D. Liau & Jeffrey J. Babon

  2. Department of Medical Biology, University of Melbourne, Melbourne, VIC, Australia

    Nicholas P. D. Liau & Jeffrey J. Babon

Authors
  1. Nicholas P. D. Liau
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  2. Jeffrey J. Babon
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Corresponding author

Correspondence to Jeffrey J. Babon .

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Editors and Affiliations

  1. Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, Victoria, Australia

    Brendan J. Jenkins

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Liau, N.P.D., Babon, J.J. (2018). Expression and Purification of JAK1 and SOCS1 for Structural and Biochemical Studies. In: Jenkins, B. (eds) Inflammation and Cancer. Methods in Molecular Biology, vol 1725. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7568-6_21

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  • DOI: https://doi.org/10.1007/978-1-4939-7568-6_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7567-9

  • Online ISBN: 978-1-4939-7568-6

  • eBook Packages: Springer Protocols

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