Use of High Performance Liquid Chromatography in Characterizing Nucleotide Binding Sites and Antigenic Determinants in cAMP-Dependent Protein Kinase

  • Susan Taylor
  • Anthony Kerlavage
  • Norman Nelson
  • Sharon Weldon
  • Mark Zoller
Part of the Experimental Biology and Medicine book series (EBAM, volume 3)


cAMP-dependent protein kinase is a multisubunit protein whose activity and quaternary structure are both modulated by cAMP (1,2). In its native holoenzyme form, the enzyme is an inactive tetramer containing two regulatory (R) and two catalytic (C) subunits. In the presence of cAMP, the holoenzyme dissociates into a regulatory subunit dimer (R2) and two monomeric catalytic subunits with each R2 binding 4 molecules of cAMP (3,4). Because of the multiple aggregation states that serve a regulatory role and because of several substrate and modulator binding sites, this enzyme provides an excellent model for investigating protein-protein interactions, nucleotide-protein interactions, and enzyme regulation. We have utilized high performance liquid chromatography (HPLC) in conjunction with solid phase sequencing to characterize a variety of specific sites on both subunits of the kinase purified from porcine cardiac muscle.


High Performance Liquid Chromato Antigenic Site Limited Proteolysis High Performance Liquid Chromato High Performance Liquid Chromato 
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  1. 1.
    Beavo, J.A., Bechtel, P.J., and Krebs, E. G. (1975) Adv. Cyclic Nuc. Res. 5, 241–251.Google Scholar
  2. 2.
    Rosen, O.M., Erlichman, J., and Rubin, C.S. (1975) Ado. Cyclic Nuc. Res. 5, 253–263.Google Scholar
  3. 3.
    Corbin, J.D., Sugden, P.H., West, L., Flockhart, D.A., Lincoln, T.M., and McCarthy, D. (1978)J. Biol. Chem. 253 3997–4003.PubMedGoogle Scholar
  4. 4.
    Builder, S.E., Beavo, J.A., and Krebs, E.G. (1980) J. Biol. Chem. 255, 2350–2354.PubMedGoogle Scholar
  5. 5.
    Hofmann, F., Beavo, J.A., Bechtel, P.J., and Krebs, E.G. (1975) J. Biol. Chem. 250, 7795–7801.PubMedGoogle Scholar
  6. 6.
    Zoller, M.J., Kerlavage, A.R., and Taylor, S.S. (1979) J. Biol, Chem. 254, 2408–2412.Google Scholar
  7. 7.
    Granot, J., Kondo, H., Armstrong, R.N., Mildvan, A.S., and Kaiser, E.T. (1979) Biochemistry 18, 2339–2345.PubMedCrossRefGoogle Scholar
  8. 8.
    Pal, K., Wechter, W.J., and Colman, R.F. (1975) J. Biol. Chem. 250, 8140–8147.PubMedGoogle Scholar
  9. 9.
    Zoller, M.J. and Taylor, S. S. (1979) J. Biol. Chem. 254, 8363–8368.PubMedGoogle Scholar
  10. 10.
    Zoller, M.J., Nelson, N.C., and Taylor, S.S. (1981)J. Biol. Chem. 256, in press.Google Scholar
  11. 11.
    Titani, K., Shoji, S., Ericsson, L.H., Demaille, J.G., Walsh, K., Neurath, H., Fischer, E.H., Takio, K., Smith, S.B., and Krebs, E.G. (1981)in Cold Spring Harbor Symposium on Protein PhosphorylationVol. 8, Book A, 19–32.Google Scholar
  12. 12.
    Nelson, N.C. and Taylor, S.S. (1981) J. Biol. Chem. 256, 3743–3750.PubMedGoogle Scholar
  13. 13.
    Potter, R.L. and Taylor, S.S. (1979) J. Biol. Chem. 254, 9000–9005.PubMedGoogle Scholar
  14. 14.
    Kerlavage, A.R. and Taylor, S.S. (1980) J. Biol. Chem. 255, 8483–8488.PubMedGoogle Scholar
  15. 15.
    Kerlavage, A.R. and Taylor, S.S. (1981)J. Biol. Chem. in press.Google Scholar
  16. 16.
    Mumby, M. and Beavo, J.A. (1981) in Cold Spring Harbor Symposium on Protein Phosphorylation, Vol. 8, Book A, 105–124.Google Scholar

Copyright information

© The HUMANA Press Inc. 1982

Authors and Affiliations

  • Susan Taylor
    • 1
  • Anthony Kerlavage
    • 1
  • Norman Nelson
    • 1
  • Sharon Weldon
    • 1
  • Mark Zoller
    • 1
  1. 1.Dept. of ChemistryUniversity of California at San DiegoLa JollaUSA

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