Summary
Using native myosin phosphorylated on either the heavy chain or the light chain, we have isolated myosin phosphatases from extracts of vegetativeDictyostelium amoeba. Two phosphatases were resolved by DEAE-cellulose chromatography. One of these phosphatases removed phosphate from heavy chain or light chain at approximately the same rate. The other phosphatase appeared to be much more specific for phosphorylated myosin heavy chain. Although these enzymes removed phosphate from other phosphoprotein substrates such as histone or casein, they did so at a much lower rate. Both enzymes required magnesium for activity, but appeared to be independent of calcium.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adelstein, R. S. &Conti, M. A. (1975) Phosphorylation of platelet myosin increases actin-activated myosin ATPase activity.Nature, Lond. 256, 597–8.
Berlot, C. H., Spudich, J. A. &Devreotes, P. N. (1985) Chemoattractant-elicited increases in myosin phosphorylation inDictyostelium.Cell 43, 307–14.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.Analyt. Biochem. 72, 248–54.
Carboni, J. M. &Condeelis, J. S. (1985) Ligand-inducec changes in the location of actin, myosin, 95K (alphaactinin), and 120K protein in amebae ofDictyostelium discoideum.J. Cell Biol. 100, 1884–93.
Clarke, M. &Spudich, J. A. (1974) Biochemical and structural studies of actomyosin-like proteins from non-muscle cells.J. molec. Biol. 86, 209–22.
Collins, J. H. &Korn, E. D. (1980) Actin activation of Ca2+-sensitive Mg2+-ATPase activity ofAcanthamoeba myosin II is enhanced by dephosphorylation of its heavy chain.J. biol. Chem. 255, 8011–4.
Collins, J. H. &Korn, E. D. (1981) Purification and characterization of actin-activatable, Ca2+-sensitive myosin II fromAcanthamoeba.J. biol. Chem. 256, 2586–95.
Condeelis, J. (1979) Isolation of concanavalin A caps during various stages of formation and their assocation with actin and myosin.J. Cell Biol. 80, 751–8.
Hammer, J. A., III Albanesi, K. P. &Korn, E. D. (1983) Purification and characterization of a myosin I heavy chain kinase fromAcanthamoeba castellanii.J. biol. Chem. 258, 10168–75.
Kuczmarski, E. R. &Spudich, J. A. (1980) Regulation of myosin self-assembly: Phosphorylation ofDictyostelium heavy chain inhibits formation of thick filaments.Proc. natn. Acad. Sci. U.S.A. 77, 7292–6.
Kuczmarski, E. R., Pagone, J. &Parysek, L. M. (1985)Dictyostelium myosin: localization of the heavy chain phosphorylation site.J. Cell Biol. 101, 159a.
Kuczmarski, E. R. (1986) Partial purification of two myosin heavy chain kinases fromDictyostelium discoideum.J. Musc. Res. Cell Motility 7, 501–9.
Mcclure, J. A. &Korn, E. D. (1983) Purification of a protein phosphatase fromAcanthamoeba that dephosphorylates and activates myosin II.J. Biol. Chem. 258, 14570–5.
Mockrin, S. C. &Spudich, J. A. (1976) Calcium control of actin-activated myosin adenosine triphosphatase fromDictyostelium discoideum.Proc. Natn. Acad. Sci. U.S.A. 73, 2321–5.
Morgan, M., Perry, S. V. &Ottaway, J. (1976) Myosin light-chain phosphatase.Biochem. J. 157, 687–97.
Pagh, K., Maruta, H., Claviez, M. &Gerisch, G. (1984) Localization of two phosphorylation sites adjacent to a region important for polymerization on the tail ofDictyostelium myosin.EMBO J. 3, 3271–8.
Pato, M. D. &Adelstein, R. S. (1983) Purification and characterization of a multisubunit phosphatase from turkey gizzard smooth muscle.J. biol. Chem. 258, 7047–54.
Peltz, G., Kuczmarski, E. R. &Spudich, J. A. (1981)Dictyostelium myosin: Characterization of chymotryptic fragments and localization of the heavy-chain phosphorylation site.J. Cell Biol. 89, 104–8.
Perrie, W. T., Smillie, L. B. &Perry, S. V. (1973) A phosphorylated light-chain component of myosin from skeletal muscle.Biochem. J. 135, 151–64
Rahmsdorf, H. J., Malchow, D. &Gerisch, G. (1978) Cyclic AMP-induced phosphorylation inDictyostelium of a polypeptide comigrating with myosin heavy chains.FEBS Lett. 88, 322–6.
Read, S. M. &Northcote, D. H. (1981) Minimization of variation in the response to different proteins of the Coomassie blue G dye-binding assay for protein.Analyt. Biochem. 116, 53–64.
Sherry, J. M. F., Gorecka, A., Aksoy, M. O., Dabrowska, R. &Hartshorne, D. J. (1978) Roles of calcium and phosphorylation in the regulation of the activity of gizzard myosin.Biochemistry 17, 4411–8.
Spudich, J. A. (1982)Dictyostelium discoideum: Methods and perspectives for study of cell motility.Meth. Cell Biol. 25, 359–64.
Tafuri, S. &Kuczmarski, E. (1985) Myosin light chain kinase fromDictyostelium.J. Cell Biol. 101, 160a.
Werth, D. K., Haeberle, J. R. &Hathaway, D. R. (1982) Purification of a myosin phosphatase from bovine aortic smooth muscle.J. biol. Chem. 257, 7306–9.
Yerna, M. J., Aksoy, M. O., Hartshorne, D. J. &Goldman, R. D. (1978) BHK 21 myosin: Isolation, biochemical characterization, and intracellular localization.J. Cell Sci. 31, 411–29.
Yumura, S. &Fukui, Y. (1985) Reversible cyclic AMP-dependent change in distribution of myosin thick filaments inDictyostelium.Nature, Lond. 314, 194–6.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kuczmarski, E.R., Pagone, J. Myosin specific phosphatases isolated fromDictyostelium discoideum . J Muscle Res Cell Motil 7, 510–516 (1986). https://doi.org/10.1007/BF01753567
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01753567