Complete recovery of the phosphoenzyme‐forming activity of nucleoside‐diphosphate kinases after reconstitution of their subunits

FEBS Letters

Ohtsuki, Kenzo¹  Yokoyama, Minehiko¹  Uesaka, Hiroshi¹ 

[1] Department of Bacteriology, Tohoku University School of Medicine, Sendai 980, Japan

关键词: Nucleoside-diphosphate kinase;    Enzyme submit;    Reconstitution;    Phosphoenzyme formation;    (HeLa S3 cell);    NDP kinase;    nucleoside-diphosphate kinase;    DTT;    dithiothreitol;    PMSF;    phenylmethylsulphonyl fluoride;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;    FPLC;    fast protein liquid chromatography;   

DOI  :  10.1016/0014-5793(86)80998-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Two distinct subunits [α-subunit (M _r 21 000, pI 7.6) and β-subunit (M _r 19000, 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

【 授权许可】

Unknown