Expression of Protein Histidine Phosphatase in Escherichia coli, Purification, and Determination of Enzyme Activity

  • Nicole Bäumer
  • Anette Mäurer
  • Josef Krieglstein
  • Susanne Klumpp
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 365)

Abstract

A protein histidine phosphatase (PHP) from vertebrates was first identified in 2002. Here we describe the expression of that PHP in Escherichia coli and purification of the recombinant protein. In addition, a detailed protocol is provided describing determination of PHP activity in vitro. Proteins phosphorylated on histidine residues in general cannot be easily obtained. This also applies to the substrates of PHP. To circumvent that obstacle, assay conditions are introduced enabling scientists to study PHP activity using a substrate within crude homogenates of cells and tissues.

Key Words

Dephosphorylation E. coli expression phosphorylation protein histidine phosphatase purification 

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Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Nicole Bäumer
    • 1
  • Anette Mäurer
    • 1
  • Josef Krieglstein
    • 2
  • Susanne Klumpp
    • 1
  1. 1.Institut für Pharmazeutische und Medizinische ChemieWestfälische Wilhelms-UniversitätMünsterGermany
  2. 2.Institut für Pharmakologie and ToxikologiePhilipps-UniversitätMarburgGermany

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