Protein Kinase CK2: Biphasic Kinetics with Peptide Substrates

Publisher： Elsevier

ISSN： 0003-9861

Source： Archives of Biochemistry and Biophysics, Vol.325, Iss.2, 1996-01, pp. : 289-294

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Abstract

The regulatory beta-subunit of the protein kinase CK2 modulates the salt optimum for alpha-subunit activity. In the presence of salt the beta-subunit is stimulatory while in the absence of salt it is inhibitory. In the presence of 150 m M NaCl CK2 has linear kinetics (Lineweaver-Burk) for the synthetic substrate RRRDDDSDDD with an apparent K m of 60 mu M . In contrast, CK2 displayed biphasic kinetics for the peptide substrate when assayed in the absence of added NaCl. Biphasic kinetics were also obtained for other peptides but not for calsequestrin or casein. Recombinant alpha-subunit had strictly linear kinetics in the absence of added NaCl with an apparent K m of 104 mu M . Preincubation of CK2 with ATP/Mg 2+ or GTP/Mg 2+ , but not adenosine/Mg 2+ or Mg 2+ alone, resulted in kinetics that were near linear. This change in kinetics was dependent on enzyme concentration but not autophosphorylation. Under conditions of low salt CK2 displays biphasic kinetics for peptide substrates, the biphasic kinetics require the presence of the beta-subunit, and ATP/Mg 2+ binding reverses the effect.