Bovine liver phosphoamidase as a protein histidine/lysine phosphatase

A 13-kDa phosphoamidase was isolated as a single band on SDS-PAGE from bovine liver. Its Stokes' radius, sedimentation coefficient, molecular mass, and optimal pH were estimated to be 1.6 nm, 1.8 s, 13 kDa, and 6.5, respectively. The enzyme released P1 from 3-phosphohistidine, 6-phospholysine,...

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Bibliographic Details
Published in:Journal of biochemistry (Tokyo) 1999-08, Vol.126 (2), p.368-374
Main Authors: Hiraishi, H, Yokoi, F, Kumon, A
Format: Article
Language:English
Subjects:
3-phosphohistidine
6-phospholysine
amidophosphate
amino acid derivatives
Animals
Cattle
characterization
Chromatography, Agarose
Chromatography, Liquid
dephosphorylation
Dose-Response Relationship, Drug
Electrophoresis, Paper
enzyme activity
Enzyme Inhibitors - pharmacology
Ethylmaleimide - pharmacology
histidine
Histidine - metabolism
Hydrogen-Ion Concentration
hydrolases
Hydrolases - chemistry
Hydrolases - isolation & purification
Kinetics
liver
Liver - enzymology
lysine
Lysine - metabolism
molecular weight
nucleoside diphosphate kinase
Nucleoside-Diphosphate Kinase - metabolism
phosphates (esters)
phosphorus
protein histidine/lysine phosphatase
purification
release
Substrate Specificity
succinic thiokinase
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Description
Summary:A 13-kDa phosphoamidase was isolated as a single band on SDS-PAGE from bovine liver. Its Stokes' radius, sedimentation coefficient, molecular mass, and optimal pH were estimated to be 1.6 nm, 1.8 s, 13 kDa, and 6.5, respectively. The enzyme released P1 from 3-phosphohistidine, 6-phospholysine, and amidophosphate at rates of 0.9, 0.6, and 2.6 /μmol/min/mg protein, respectively. However, it did not dephosphorylate phosphocrea-tine, Nω-phosphoarginine, imidodiphosphate, or O-phosphorylated compounds including inorganic pyrophosphate. It also dephosphorylated succinic thiokinase and nucleoside diphosphate kinase autophosphorylated at His residues, indicating that it works as a protein histidine phosphatase. A thiol reagent, 30 μM N-ethylmaleimide, depressed the activity by half, while a thiol compound, 2-mercaptoethanol, protected the enzyme from heat-inactivation. Five millimolar divalent cations, such as Mg2+ and Mn2+, and 5 mM EDTA, had no effect on the activity.
ISSN:0021-924X
1756-2651
DOI:10.1093/oxfordjournals.jbchem.a022459