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Complete recovery of the phosphoenzyme‐forming activity of nucleoside‐diphosphate kinases after reconstitution of their subunits
Author(s) -
Yokoyama Minehiko,
Uesaka Hiroshi,
Ohtsuki Kenzo
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)80998-x
Subject(s) - protein subunit , kinase , fast protein liquid chromatography , chemistry , nucleoside , enzyme , biochemistry , urea , hela , in vitro , microbiology and biotechnology , biology , gene
Two distinct subunits [α‐subunit ( M r 21 000, p I 7.6) and β‐subunit ( M r 19000, p I 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 [ 32 P]‐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