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Histidine Phosphorylation pp 109–120Cite as

In Vitro Assays for Measuring Protein Histidine Phosphatase Activity

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 2077))

Abstract

In order to obtain a detailed kinetic characterization, identify inhibitors, and elucidate the biological roles of an enzyme, it is advantageous to have a facile, sensitive enzyme assay protocol. Here we present a brief overview of the techniques available to monitor histidine phosphatase activity and provide protocols for measuring the activity and inhibition of PHPT1 in vitro using the fluorescent probe 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP). This assay uses small quantities of commercially available materials, making its use feasible for most laboratories.

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Acknowledgments

This work was supported by a Teva Pharmaceuticals Mark A. Goshko Memorial Grant award (56426-TEV) and an NSF award (CHE 1308766) to A.M.B.

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Authors and Affiliations

  1. Department of Medicinal Chemistry, University of Utah, Salt Lake City, UT, USA

    Brandon S. McCullough & Amy M. Barrios

Authors
  1. Brandon S. McCullough
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  2. Amy M. Barrios
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Corresponding author

Correspondence to Amy M. Barrios .

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Editors and Affiliations

  1. Department of Biochemistry, Centre for Proteome Research Institute of Integrative Biology, University of Liverpool, Liverpool, UK

    Claire E. Eyers

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McCullough, B.S., Barrios, A.M. (2020). In Vitro Assays for Measuring Protein Histidine Phosphatase Activity. In: Eyers, C. (eds) Histidine Phosphorylation. Methods in Molecular Biology, vol 2077. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9884-5_8

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  • DOI: https://doi.org/10.1007/978-1-4939-9884-5_8

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9883-8

  • Online ISBN: 978-1-4939-9884-5

  • eBook Packages: Springer Protocols

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