Nucleoside-triphosphatase and hydrolysis of thiamin triphosphate in Escherichia coli.
A membrane-bound nonspecific triphosphatase of E. coli was solubilized and purified to a homogeneous SDS-acrylamide gel electrophoresis band. It was found to be a single polypeptide of 16 kDa requiring no Mg2+, with an optimal pH at 6.5. The substrate specificity was broad and a nonspecific Mg2+-independent ribonucleoside-triphosphatase (NTPase) activity was expressed together with thiamin-triphosphatase activity. The molecular size and characteristics were clearly different from the known NTPase (EC 3.6.1.15). Using the purified thiamin-triphosphatase II, ATP:thiamin-diphosphate phosphoryl transferase (EC 2.7.4.15) activity was demonstrated with an optimal pH of approx. 5.3. Considering its kinetic parameters and other characteristics, however, the thiamin triphosphate synthesizing activity was not thought to take part in cellular thiamin triphosphate synthesis. The possibility that thiamin-triphosphatase II plays a part in the hydrolysis of thiamin triphosphate to control its cellular level is suggested.
D002462:Cell Membrane / Q000201:enzymology; D002845:Chromatography; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification / Q000378:metabolism*; D010770:Phosphotransferases / Q000302:isolation & purification / Q000378:metabolism; D017856:Phosphotransferases (Phosphate Group Acceptor); D013379:Substrate Specificity; D013837:Thiamin-Triphosphatase / Q000302:isolation & purification / Q000378:metabolism*; D013831:Thiamine / Q000031:analogs & derivatives*; D013838:Thiamine Triphosphate / Q000378:metabolism*
D013838:Thiamine Triphosphate; D010770:Phosphotransferases; D017856:Phosphotransferases (Phosphate Group Acceptor); C022733:thiamin-diphosphate kinase; D010744:Phosphoric Monoester Hydrolases; D013837:Thiamin-Triphosphatase; D008274:Magnesium; D013831:Thiamine