Publications

• Protein histidine phosphatase: a novel enzyme with potency for neuronal signaling.

  Klumpp S., Hermesmeier J., Selke D., Bechmann G., van den Brulle J., Weidner G., Scharm B., Guessow D., Baumeister R., Kellner R., Krieglstein J.

  The importance of reversible phosphorylation for neuronal signaling and cell survival is well recognized. Knowledge in vertebrates, however, is so far limited to O-phosphates from serine, threonine, and tyrosine. The authors describe an enzyme acting on N-phosphates. It is the first protein histid ... >> More

  The importance of reversible phosphorylation for neuronal signaling and cell survival is well recognized. Knowledge in vertebrates, however, is so far limited to O-phosphates from serine, threonine, and tyrosine. The authors describe an enzyme acting on N-phosphates. It is the first protein histidine phosphatase identified in vertebrates. This histidine phosphatase is ubiquitously expressed in mammalian tissues including brain. Characterization and sequencing showed a yet unknown protein with no similarity to other phosphatases. In Caenorhabditis elegans, the homolog of this histidine phosphatase was exclusively expressed in neurons, suggesting a distinct role of reversible histidine phosphorylation in neuronal functions. << Less

  J. Cereb. Blood Flow Metab. 22:1420-1424(2002) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Expression of protein histidine phosphatase in Escherichia coli, purification, and determination of enzyme activity.

  Baumer N., Maurer A., Krieglstein J., Klumpp S.

  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 ... >> More

  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. << Less

  Methods Mol Biol 365:247-260(2007) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Identification and characterization of a mammalian 14-kDa phosphohistidine phosphatase.

  Ek P., Pettersson G., Ek B., Gong F., Li J.-P., Zetterqvist O.

  Protein histidine phosphorylation in eukaryotes has been sparsely studied compared to protein serine/threonine and tyrosine phosphorylation. In an attempt to rectify this by probing porcine liver cytosol with the phosphohistidine-containing peptide succinyl-Ala-His(P)-Pro-Phe-p-nitroanilide (phosp ... >> More

  Protein histidine phosphorylation in eukaryotes has been sparsely studied compared to protein serine/threonine and tyrosine phosphorylation. In an attempt to rectify this by probing porcine liver cytosol with the phosphohistidine-containing peptide succinyl-Ala-His(P)-Pro-Phe-p-nitroanilide (phosphopeptide I), we observed a phosphatase activity that was insensitive towards okadaic acid and EDTA. This suggested the existence of a phosphohistidine phosphatase different from protein phosphatase 1, 2A and 2C. A 1000-fold purification to apparent homogeneity gave a 14-kDa phosphatase with a specific activity of 3 micro mol.min-1.mg-1 at pH 7.5 with 7 micro m phosphopeptide I as substrate. Partial amino-acid sequence determination of the purified porcine enzyme by MS revealed similarity with a human sequence representing a human chromosome 9 gene of hitherto unknown function. Molecular cloning from a human embryonic kidney cell cDNA-library followed by expression and purification, yielded a protein with a molecular mass of 13 700 Da, and an EDTA-insensitive phosphohistidine phosphatase activity of 9 micro mol.min-1.mg-1 towards phosphopeptide I. No detectable activity was obtained towards a set of phosphoserine-, phosphothreonine-, and phosphotyrosine peptides. Northern blot analysis indicated that the human phosphohistidine phosphatase mRNA was present preferentially in heart and skeletal muscle. These results provide a new tool for studying eukaryotic histidine phosphorylation/dephosphorylation. << Less

  Eur. J. Biochem. 269:5016-5023(2002) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Phosphohistidine phosphatase 1 (PHPT1) also dephosphorylates phospholysine of chemically phosphorylated histone H1 and polylysine.

  Ek P., Ek B., Zetterqvist O.

  <h4>Background</h4>Phosphohistidine phosphatase 1 (PHPT1), also named protein histidine phosphatase (PHP), is a eukaryotic enzyme dephosphorylating proteins and peptides that are phosphorylated on a histidine residue. A preliminary finding that histone H1, which lacks histidine, was phosphorylated ... >> More

  <h4>Background</h4>Phosphohistidine phosphatase 1 (PHPT1), also named protein histidine phosphatase (PHP), is a eukaryotic enzyme dephosphorylating proteins and peptides that are phosphorylated on a histidine residue. A preliminary finding that histone H1, which lacks histidine, was phosphorylated by phosphoramidate and dephosphorylated by PHPT1 prompted the present investigation.<h4>Methods</h4>Histone H1 and polylysine were phosphorylated at a low concentration (3.9 mM) of phosphoramidate. Their dephosphorylation by recombinant human PHPT1 was investigated by using a DEAE-Sepharose spin column technique earlier developed by us for studies on basic phosphoproteins and phosphopeptides. Determination of protein-bound, acid-labile phosphate was performed by a malachite green method. Mass spectrometry (MS) was used to investigate the occurrence of N-ε-phospholysine residues in a phosphorylated histone H1.2 preparation, and to measure the activity of PHPT1 against free N-ω-phosphoarginine.<h4>Results</h4>Histone H1.2, which lacks histidine, was phosphorylated by phosphoramidate on several lysine residues, as shown by MS. PHPT1 was shown to dephosphorylate phosphohistone H1 at a rate similar to that previously described for the dephosphorylation of phosphohistidine-containing peptides. In addition, phosphopolylysine was an equally good substrate for PHPT1. However, no dephosphorylation of free phosphoarginine by PHPT1 could be detected.<h4>Conclusion</h4>The finding that PHPT1 can dephosphorylate phospholysine in chemically phosphorylated histone H1 and polylysine demonstrates a broader specificity for this enzyme than known so far. << Less

  Ups. J. Med. Sci. 120:20-27(2015) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Chemical phosphorylation of histidine-containing peptides based on the sequence of histone H4 and their dephosphorylation by protein histidine phosphatase.

  Attwood P.V., Ludwig K., Bergander K., Besant P.G., Adina-Zada A., Krieglstein J., Klumpp S.

  Using peptides based on the amino acid sequences surrounding the two histidine residues in histone H4, we have investigated the kinetics of the phosphorylation and dephosphorylation reactions of their histidine residues, when reacted with potassium phosphoramidate, by (1)H NMR. We have been able t ... >> More

  Using peptides based on the amino acid sequences surrounding the two histidine residues in histone H4, we have investigated the kinetics of the phosphorylation and dephosphorylation reactions of their histidine residues, when reacted with potassium phosphoramidate, by (1)H NMR. We have been able to estimate rate constants for the reactions and have shown that there are differences in the kinetics between the two peptides. The kinetics of hydrolysis of phosphoramidate was measured by (31)P NMR and protein histidine phosphatase (PHP) was shown to catalyse the reaction. We have shown that the dephosphorylation of the phosphohistidine of the phosphopeptides is catalysed by PHP. In terms of substrate specificity, there is a small preference for 1-phosphohistidine compared to 3-phosphohistidine, although the rate accelerations for hydrolysis induced by the enzyme were 1100- and 33,333-fold, respectively. The kinetics of both the phosphorylation and dephosphorylation reactions depend on the amino acid sequence surrounding the histidine. PHP shows greater substrate specificity for the peptide whose sequence is similar to that around histidine 18 of histone H4. PHP was unable to catalyse the dephosphorylation of histone H4 that had been phosphorylated with a histone H4 histidine kinase. << Less

  Biochim. Biophys. Acta 1804:199-205(2010) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.