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Assessment of Cytokine-Modulated Proteasome Activity

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Cytokine Bioassays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1172))

Abstract

This chapter presents two methods for assessment of proteasome function. The first is a modification of the standard fluorogenic peptide cleavage assay which takes into account the effect of ATP on proteasome activity. This method is described in both its macro and high throughput micro-assay forms. The second is the Proteasome Constitutive Immuno-Subunit (active site) ELISA or ProCISE method. ProCISE is a modification of active site directed probe analysis and allows for convenient differentiation between active constitutive and immuno-subunits. While the utility of measuring proteasome activity and its relationship to cytokine action and inflammation are clear, the assessment and interpretation is not always straightforward. Therefore, we also discuss the pitfalls of the standard fluorogenic assay, particularly in the interpretation of results obtained, and the advantages of the newer, ProCISE assay.

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Author information

Authors and Affiliations

  1. Onyx Pharmaceuticals, 249 Grand Avenue South, San Francisco, CA, 94080, USA

    Christopher J. Kirk Ph.D.

  2. Cardiac Metabolism Laboratory, Feinstein Institute for Medical Research, Manhasset, NY, 11030, USA

    Saul R. Powell Ph.D., F.A.H.A.

  3. Molecular Medicine and Science Education, Hofstra North Shore-LIJ Medical School, Hempstead, NY, 11549, USA

    Saul R. Powell Ph.D., F.A.H.A.

  4. Center for Heart and Lung Research, Feinstein Institute for Medical Research, 350 Community Drive, Manhasset, NY, 11030-816, USA

    Edmund J. Miller Ph.D., C.Chem., F.R.S.C.

  5. Medicine and Molecular Medicine, Hofstra North Shore-LIJ Medical School, Hempstead, NY, 11030-816, 11549, USA

    Edmund J. Miller Ph.D., C.Chem., F.R.S.C.

Authors
  1. Christopher J. Kirk Ph.D.
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  2. Saul R. Powell Ph.D., F.A.H.A.
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  3. Edmund J. Miller Ph.D., C.Chem., F.R.S.C.
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Corresponding author

Correspondence to Edmund J. Miller Ph.D., C.Chem., F.R.S.C. .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, St. John's University, New York, New York, USA

    Ivana Vancurova

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Kirk, C.J., Powell, S.R., Miller, E.J. (2014). Assessment of Cytokine-Modulated Proteasome Activity. In: Vancurova, I. (eds) Cytokine Bioassays. Methods in Molecular Biology, vol 1172. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0928-5_13

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  • DOI: https://doi.org/10.1007/978-1-4939-0928-5_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0927-8

  • Online ISBN: 978-1-4939-0928-5

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