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Using the activated cGMP-dependent protein kinase in the presence of the phosphorylatable peptide [[Ala34]histone H2B-(29-35)], we found that lin-benzoadenosine 5'-diphosphate (lin-benzo-ADP) was a competitive inhibitor of the enzyme with respect to ATP with a Ki (22 microM) similar to the Kd (20 microM) determined by fluorescence polarization titrations. The Kd for lin-benzo-ADP determined in the absence of the phosphorylatable peptide, however, was only 12 microM. ADP bound with lower affinity (Ki = 169 microM; Kd = 114 microM). With [Ala34]histone H2B-(29-35) as phosphoryl acceptor, the Km for lin-benzo-ATP was 29 microM, and that for ATP was 32 microM. The Vmax with lin-benzo-ATP, however, was only 0.06% of that with ATP as substrate [0.00623 +/- 0.00035 vs. 11.1 +/- 0.17 mumol (min.mg)-1]. Binding of lin-benzo-ADP to the kinase was dependent upon a divalent cation. Fluorescence polarization revealed that Mg2+, Mn2+, Co2+, Ni2+, Ca2+, Sr2+, and Ba2+ supported nucleotide binding to the enzyme; Ca2+, Sr2+, and Ba2+, however, did not support any measurable phosphotransferase activity. The rank order of metal ion effectiveness in mediating phosphotransferase activity was Mg2+ greater than Ni2+ greater than Co2+ greater than Mn2+. Although these results were similar to those observed with the cAMP-dependent protein kinase [Hartl, F. T., Roskoski, R., Jr., Rosendahl, M. S., & Leonard, N. J. (1983) Biochemistry 22, 2347], major differences in the Vmax with lin-benzo-ATP as substrate and the effect of peptide substrates on nucleotide (both lin-benzo-ADP and ADP) binding were observed.
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
May 1983,
Biochemistry,
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
August 1984,
Biochemistry,
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
April 1979,
Biochimica et biophysica acta,
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
January 1979,
Current topics in cellular regulation,
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
April 1977,
The Journal of biological chemistry,
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
June 1978,
Journal of cyclic nucleotide research,
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
January 1999,
Zeitschrift fur Naturforschung. C, Journal of biosciences,
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
August 2005,
Journal of immunology (Baltimore, Md. : 1950),
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
June 1979,
The Journal of biological chemistry,
D Bhatnagar, and
D B Glass, and
R Roskoski, and
R A Lessor, and
N J Leonard
December 2011,
Cancer,
