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. 1990 Jun 1;268(2):367–370. doi: 10.1042/bj2680367

Kinase activity associated with caldesmon is Ca2+/calmodulin-dependent kinase II.

G C Scott-Woo 1, C Sutherland 1, M P Walsh 1
PMCID: PMC1131441  PMID: 2163610

Abstract

The relationship of the kinase which co-purifies with caldesmon to Ca2+/calmodulin-dependent protein kinase II (CaM-kinase II) was investigated by studying the phosphorylation of bovine brain synapsin I, as well-characterized substrate of CaM-kinase II. Synapsin I is a very good substrate (Km = 90 nM) of the co-purifying kinase, which phosphorylates two sites in synapsin I, both of which are distinct from the single site phosphorylated by cyclic-AMP-dependent protein kinase. Phosphorylation of synapsin I is Ca2(+)- and calmodulin-dependent: half-maximal activation occurs at 0.13 microM-Ca2+ and maximal activity at 0.4 microM-Ca2+. Phosphorylation of the co-purifying kinase slightly enhances the rate, but does not alter the stoichiometry, of subsequent synapsin I phosphorylation; it does, however, circumvent the requirement for Ca2+ and calmodulin. The properties of this kinase therefore closely resemble those of CaM-kinase II, and we conclude that it is probably a smooth-muscle isoenzyme of CaM-kinase II.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abougou J. C., Hagiwara M., Hachiya T., Terasawa M., Hidaka H., Hartshorne D. J. Phosphorylation of caldesmon. FEBS Lett. 1989 Nov 6;257(2):408–410. doi: 10.1016/0014-5793(89)81583-2. [DOI] [PubMed] [Google Scholar]
  2. Adam L. P., Haeberle J. R., Hathaway D. R. Phosphorylation of caldesmon in arterial smooth muscle. J Biol Chem. 1989 May 5;264(13):7698–7703. [PubMed] [Google Scholar]
  3. Bryan J., Imai M., Lee R., Moore P., Cook R. G., Lin W. G. Cloning and expression of a smooth muscle caldesmon. J Biol Chem. 1989 Aug 15;264(23):13873–13879. [PubMed] [Google Scholar]
  4. Clark T., Ngai P. K., Sutherland C., Gröschel-Stewart U., Walsh M. P. Vascular smooth muscle caldesmon. J Biol Chem. 1986 Jun 15;261(17):8028–8035. [PubMed] [Google Scholar]
  5. Colbran R. J., Schworer C. M., Hashimoto Y., Fong Y. L., Rich D. P., Smith M. K., Soderling T. R. Calcium/calmodulin-dependent protein kinase II. Biochem J. 1989 Mar 1;258(2):313–325. doi: 10.1042/bj2580313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Czernik A. J., Pang D. T., Greengard P. Amino acid sequences surrounding the cAMP-dependent and calcium/calmodulin-dependent phosphorylation sites in rat and bovine synapsin I. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7518–7522. doi: 10.1073/pnas.84.21.7518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Demaille J. G., Peters K. A., Fischer E. H. Isolation and properties of the rabbit skeletal muscle protein inhibitor of adenosine 3',5'-monophosphate dependent protein kinases. Biochemistry. 1977 Jul 12;16(14):3080–3086. doi: 10.1021/bi00633a006. [DOI] [PubMed] [Google Scholar]
  8. Fabiato A. Myoplasmic free calcium concentration reached during the twitch of an intact isolated cardiac cell and during calcium-induced release of calcium from the sarcoplasmic reticulum of a skinned cardiac cell from the adult rat or rabbit ventricle. J Gen Physiol. 1981 Nov;78(5):457–497. doi: 10.1085/jgp.78.5.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Huttner W. B., DeGennaro L. J., Greengard P. Differential phosphorylation of multiple sites in purified protein I by cyclic AMP-dependent and calcium-dependent protein kinases. J Biol Chem. 1981 Feb 10;256(3):1482–1488. [PubMed] [Google Scholar]
  10. Nairn A. C., Hemmings H. C., Jr, Greengard P. Protein kinases in the brain. Annu Rev Biochem. 1985;54:931–976. doi: 10.1146/annurev.bi.54.070185.004435. [DOI] [PubMed] [Google Scholar]
  11. Ngai P. K., Walsh M. P. Detection of caldesmon in muscle and non-muscle tissues of the chicken using polyclonal antibodies. Biochem Biophys Res Commun. 1985 Mar 15;127(2):533–539. doi: 10.1016/s0006-291x(85)80192-3. [DOI] [PubMed] [Google Scholar]
  12. Ngai P. K., Walsh M. P. Inhibition of smooth muscle actin-activated myosin Mg2+-ATPase activity by caldesmon. J Biol Chem. 1984 Nov 25;259(22):13656–13659. [PubMed] [Google Scholar]
  13. Ngai P. K., Walsh M. P. The effects of phosphorylation of smooth-muscle caldesmon. Biochem J. 1987 Jun 1;244(2):417–425. doi: 10.1042/bj2440417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Scott-Woo G. C., Walsh M. P. Autophosphorylation of smooth-muscle caldesmon. Biochem J. 1988 Jun 1;252(2):463–472. doi: 10.1042/bj2520463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Scott-Woo G. C., Walsh M. P. Characterization of the autophosphorylation of chicken gizzard caldesmon. Biochem J. 1988 Nov 1;255(3):817–824. doi: 10.1042/bj2550817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sobue K., Muramoto Y., Fujita M., Kakiuchi S. Purification of a calmodulin-binding protein from chicken gizzard that interacts with F-actin. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5652–5655. doi: 10.1073/pnas.78.9.5652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ueda T., Greengard P. Adenosine 3':5'-monophosphate-regulated phosphoprotein system of neuronal membranes. I. Solubilization, purification, and some properties of an endogenous phosphoprotein. J Biol Chem. 1977 Jul 25;252(14):5155–5163. [PubMed] [Google Scholar]
  18. Walsh M. P. Caldesmon, a major actin- and calmodulin-binding protein of smooth muscle. Prog Clin Biol Res. 1987;245:119–141. [PubMed] [Google Scholar]
  19. Walsh M. P., Hinkins S., Dabrowska R., Hartshorne D. J. Smooth muscle myosin light chain kinase. Methods Enzymol. 1983;99:279–288. doi: 10.1016/0076-6879(83)99063-8. [DOI] [PubMed] [Google Scholar]
  20. Walsh M. P., Valentine K. A., Ngai P. K., Carruthers C. A., Hollenberg M. D. Ca2+-dependent hydrophobic-interaction chromatography. Isolation of a novel Ca2+-binding protein and protein kinase C from bovine brain. Biochem J. 1984 Nov 15;224(1):117–127. doi: 10.1042/bj2240117. [DOI] [PMC free article] [PubMed] [Google Scholar]

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