title: The role of histidine-118 of inorganic pyrophosphatase from thermophilic bacterium PS-3.
abstract: Treatment of the inorganic pyrophosphatase from thermophilic bacterium PS-3 with diethyl pyrocarbonate resulted in the almost complete loss of its activity, which followed pseudo-first-order kinetics. The presence of Mg2+ prevented the inactivation. Enzyme inactivated with diethyl pyrocarbonate was re-activated by hydroxylamine. The inactivation parallelled the amount of modified histidine residue, and a plot of the activity remaining against the amount of modified histidine residue suggested that the modification of one of two histidine residues totally inactivated the enzyme. The site involved was found to be located in a single lysyl endopeptidase-digest peptide derived from the ethoxy[14C]carbonylated enzyme. Amino acid analysis and sequence analysis of the peptide revealed that it comprised residues 96-119 of the inorganic pyrophosphatase from thermophilic bacterium PS-3. These results, when compared with those reported for the Escherichia coli and yeast enzymes, imply that His-118 of the inorganic pyrophosphatase from thermophilic bacterium PS-3 is located near the Mg(2+)-binding site and thus affects the binding of Mg2+.
authors.affiliation: Institute of High Polymer Research, Faculty of Textile Science and Technology, Shinshu University, Nagano, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Hirano
authors.forename: T
authors.initials: T
authors.lastname: Ichiba
authors.forename: A
authors.initials: A
authors.lastname: Hachimori
mesh_terms: D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D004047:Diethyl Pyrocarbonate / Q000737:chemistry; D006639:Histidine / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D011755:Pyrophosphatases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013824:Thermus / Q000201:enzymology*
chemical_list: D000596:Amino Acids; D006639:Histidine; D011755:Pyrophosphatases; D008274:Magnesium; D004047:Diethyl Pyrocarbonate