Abstract
The catalytic subunits of adenosine 3′:5′ monophosphate-dependent protein kinase (ATP:protein transferase, E.C. 2.7.1.1.37) from the soluble and membrane fractions of swine kidney were purified to homogeneity by a new procedure and their structural, kinetic and immunological properties were compared. The specific activities of the purified enzymes were 2.35 and 2.6 µmol/min/mg of protein, with histone as the substrate. Both preparations contained a single polypeptide chain, and only one band was observed upon polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The molecular weight of both enzymes determined by gel electrophoresis was 42 000 ± 1000, and sedimentation equilibrium yielded a value of 41000 ± 800. Analysis by sedimentation velocity showed the presence of a single peak with and S20,w of 3.1 ± 0.2 for each preparation.
The amino acid compositions are very similar, and each enzyme contains about one residue of cysteine which is essential for enzymatic activity. ATP and Mg2+ protect both enzymes from inhibition by thiol specific reagents to the same extent. The catalytic subunits have similar apparent K m's for protein substrates. The enzymes exhibit single completely confluent precipitin lines when examined by immunodiffusion and the particulate catalytic subunit competitively displaces the soluble 125I-catalytic subunit in homologous radioimmunoassays. The soluble and particulate 125I-catalytic subunits bind to the regulatory subunits in the washed plasma membranes with attendent loss of kinase activity, which could be reversed by cyclic AMP. The results of experiments with kidney cortex slices treated with parathyroid hormone, epinephrine or dibutyryl cyclic AMP showed the translocation of phosphotransferase activity from the cytosol to the particulate membrane fraction. Taken collectively, these observations suggest that only one form of the catalytic subunit of cyclic AMP-dependent protein kinase is present in swine kidney, and that it may exchange between the cytosol and membrane fractions in response to specific physiological signals.
Similar content being viewed by others
References
Krebs, E. G. and Beavo, J. A., 1979. Ann. Rev. Biochem. 48: 923–959.
Rubin, C. S. and Rosen, O. M., 1975. Ann. Rev. Biochem. 44: 831–887.
Nimmo, H. G. and Cohen, P., 1977. Adv. Cyclic Nucleotide Res. 8: 145–266.
Corbin, J. D., Keely, S. L. and Park, C. R., 1975. J. Biol. Chem. 250: 218–225.
Bechtel, P. J., Beavo, J. A. and Krebs, E. G., 1977. J. Biol. Chem. 252: 2691–2697.
Rubin, C. S., Erlichman, J. and Rosen, O. M., 1972. J. Biol. Chem. 247: 36–43.
Abou-Issa, H., Kratowich, N. and Mendicino, J., 1974. Eur. J. Biochem. 42: 461–473.
Hofmann, F., Beavo, J. A., Bechtel, P. J. and Krebs, E. G., 1975. J. Biol. Chem. 250: 7795–7801.
Reiman, E. M., Walsh, D. A. and Krebs, E. G., 1971. J. Biol. Chem. 246: 1986–1995.
Erlichman, J., Rubin, C. S. and Rosen, O. M., 1973. J. Biol. Chem. 248: 7607–7609.
Miyamoto, E., Petzold, G. L., Koo, J. F. and Greengard, P., 1973. J. Biol. Chem. 248: 179–189.
Tao, M. and Hackett, P., 1973. J. Biol. Chem. 248: 5324–5330.
Garber, D. L., First, N. L. and Lardy, H. A., 1973. J. Biol. Chem. 248: 375–383.
Maeno, H., Johnson, E. M. and Greengard, P., 1971. J. Biol. Chem. 246: 134–141.
Uno, L., Ueda, T. and Greengard, P., 1977. J. Biol. Chem. 252: 5164–5174.
Rubin, C. S., Aldao, R., Sarkar, D., Erlichman, J. and Fleisher, N., 1979. J. Biol. Chem. 254: 3797–3805.
Corbin, J. D., Sugden, P. H., Lincoln, T. M. and Keely, S. L., 1977. J. Biol. Chem. 252: 3854–3861.
Uno, I., Ueda, T. and Greengard, P., 1976. J. Biol. Chem. 251: 2192–2195.
Mendicino, J., Abou-Issa, H., Medicus, R. and Kratowich, N., 1975. Methods Enzymol. (Ed. Wood, W. A.) Vol. 42, pp. 375–397, Academic Press, New York.
Abou-Issa, H. and Mendicino, J., 1973. J. Biol. Chem. 248: 685–696.
Gilman, A. G., 1970. Proc. Natl. Acad. Sci. (USA) 67: 305–312.
Hunter, W. M., 1967. in Handbook of Experimental Immunology (Ed. Weir, D. M.) pp. 608–654, Blackwell Scientific PublicationsEdinburgh.
Utsumi, S. and Karush, F., 1965. Biochemistry 4: 1766–1779.
Dus, K. M., Litchfield, W. J., Hippenmeyer, Bumpus, J. A., Obidoa, O., Spitsberg, V. and Jefcoate, C. R., 1980. Eur. J. Biochem. 111: 307–314.
Greenwood, F. C., Hunter, W. M. and Glover, S. J., 1963. Biochem. J. 89: 114–123.
Hewitt, J., Pillion, D. J. and Leibach, F. H., 1974. Biochim. Biophys. Acta. 363: 267–276.
Pillion, D. J., Leibach, F. H., Von Tersch, F. and Mendicino, J., 1976. Biochim. Biophys. Acta. 419: 104–111.
Abou-Issa, H., Mendicino, J., Leibach, F. H. and Pillion, D. J., 1975. FEBS Lett. 50: 121–124.
Fitzpatrick, D. F., Davenport, G. R., Forte, L. and Landon, E. J., 1969. J. Biol. Chem. 244: 3561–3569.
Mendicino, J. and Kratowich, N., 1972. J. Biol. Chem. 247: 6643–6650.
Martin, R. G. and Ames, B. N., 1961. J. Biol. Chem. 236: 1372–1379.
Weber, K. and Osborn, M., 1969. J. Biol. Chem. 244: 2406–2412.
Cuatrecasas, P., 1970. J. Biol. Chem. 245: 3059–3065.
Travis, J. and Parnnell, R., 1973. Clin. Chim. Acta 49: 49–52.
Travis, J., Bowen, J., Tewksbury, D., Johnson, D. and Parnnell, R., 1976. Biochem. J. 157: 301–306.
Glynn, I. M. and Chappell, J. B., 1964. Biochem. J. 90: 147–149.
Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J., 1951. J. Biol. Chem. 193: 265–275.
Bradford, M. M., 1976. Anal. Biochem. 72: 248–254.
Mendicino, J., Leibach, F. H. and Reddy, S., 1978. Biochemistry 17: 4662–4668.
Hui, C., Drummond, M. and Drummond, G. I., 1976. Arch. Biochem. Biophys. 173: 415–427.
St. Louis, P. J. and Sulakhe, P. V., 1979. Arch. Biochem. Biophys. 198: 227–240.
Walter, U., Lohman, S. M., Sieghart, W. and Greengard, P., 1979. J. Biol. Chem. 254: 12235–12239.
Jones, L. R., Maddock, S. W. and Hathaway, D. R., 1981. Biochim. Biophys. Acta 641: 242–253.
Dreyfuss, G., Schwartz, K. J. and Blout, E. R., 1978. Proc. Natl. Acad. Sci. (USA) 75: 5926–5930.
Rubin, C. S., 1979. J. Biol. Chem. 254: 12439–12449.
Chang, K. W. and Cautrecasas, P., 1974. J. Biol. Chem. 249: 3170–3180.
Krichberger, M. A., Tada, M. and Katz, A. M., 1974. J. Biol. Chem. 249: 6166–6172.
Mallette, L. E., Exton, J. H. and Park, C. R., 1969. J. Biol. Chem. 244: 5713–5723.
Lambert, M., Camus, J. and Christophe, J., 1973. Biochem. Biophys. Res. Commun. 52: 935–942.
Majumdar, G. C. and Turkington, R. W., 1972. J. Biol. Chem. 247: 7207–7217.
Labrie, F., Lemaire, S., Poirier, G., Pelletier, G. and Boucher, R., 1971. J. Biol. Chem. 246: 7311–7317.
Rubin, C. S., 1975. J. Biol. Chem. 250: 9044–9052.
Keely, S. L., Corbin, J. D. and Park, C. R., 1975. Proc. Natl. Acad. Sci. (USA) 72: 1501–1504.
Szoka, F. C. and Ettinger, M. J., 1978. J. Biol. Chem. 253: 6536–6543.
Morel, F., 1981. Am. J. Physiol. 240: F159–164.
Fishman, P. H. and Atkkan, E. F., 1980. J. Membrane Biol. 54: 51–60.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Muniyappa, K., Leibach, F.H. & Mendicino, J. Characterization and comparison of the soluble and membrane-bound cyclic AMP-dependent protein kinase from swine kidney. Mol Cell Biochem 50, 157–171 (1983). https://doi.org/10.1007/BF00285641
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00285641