Complete recovery of the phosphoenzyme-forming activity of nucleoside-diphosphate kinases after reconstitution of their subunits.
Two distinct subunits [alpha-subunit (Mr 21 000, pI 7.6) and beta-subunit (Mr 19 000, pI 6.5)] of nucleoside-diphosphate (NDP) kinases highly purified from HeLa S3 cells can be separated by FPLC using a Mono P column in the presence of 6 M urea and 1% pharmalyte (pH range between 5.0 and 8.0). Comparatively high [32P]phosphate incorporation was detected when these two subunit fractions were reconstituted in vitro. Available evidence suggests that these two enzyme subunits are necessary for the formation of phosphoenzyme, which functions as an intermediate in NDP kinase action.
D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D009701:Nucleoside-Diphosphate Kinase / Q000302:isolation & purification / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D010770:Phosphotransferases / Q000378:metabolism*
D010710:Phosphates; D010750:Phosphoproteins; D010770:Phosphotransferases; D009701:Nucleoside-Diphosphate Kinase