title: Chemical phosphorylation of histidine-containing peptides based on the sequence of histone H4 and their dephosphorylation by protein histidine phosphatase.
abstract: 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.
authors.affiliation: School of Biomedical, Biomolecular and Chemical Sciences, The University of Western Australia, Crawley, WA 6009, Australia. pattwood@cyllene.uwa.edu.au
authors.forename: Paul V
authors.initials: PV
authors.lastname: Attwood
authors.forename: Katrin
authors.initials: K
authors.lastname: Ludwig
authors.forename: Klaus
authors.initials: K
authors.lastname: Bergander
authors.forename: Paul G
authors.initials: PG
authors.lastname: Besant
authors.forename: Abdussalam
authors.initials: A
authors.lastname: Adina-Zada
authors.forename: Josef
authors.initials: J
authors.lastname: Krieglstein
authors.forename: Susanne
authors.initials: S
authors.lastname: Klumpp
mesh_terms: D000577:Amides / Q000737:chemistry; D000595:Amino Acid Sequence; D006639:Histidine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000235:genetics; D000071677:Histidine Kinase; D006657:Histones / Q000737:chemistry; D007700:Kinetics; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000378:metabolism; D010756:Phosphoric Acids / Q000737:chemistry; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism
chemical_list: D000577:Amides; D006657:Histones; D010455:Peptides; D010756:Phosphoric Acids; D006639:Histidine; C011067:phosphoramidic acid; D011494:Protein Kinases; D000071677:Histidine Kinase; D010744:Phosphoric Monoester Hydrolases; C005436:phosphohistidine