title: Expression of protein histidine phosphatase in Escherichia coli, purification, and determination of enzyme activity.
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.
authors.affiliation: Institut für Pharmazeutische und Medizinische Chemie, Westfälische Wilhelms-Universität, Münster, Germany.
authors.forename: Nicole
authors.initials: N
authors.lastname: Bäumer
authors.forename: Anette
authors.initials: A
authors.lastname: Mäurer
authors.forename: Josef
authors.initials: J
authors.lastname: Krieglstein
authors.forename: Susanne
authors.initials: S
authors.lastname: Klumpp
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics*; D006639:Histidine / Q000378:metabolism*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D010766:Phosphorylation; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D006639:Histidine; D010744:Phosphoric Monoester Hydrolases