Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1994 Apr 15;299(Pt 2):489–495. doi: 10.1042/bj2990489

New isoforms of Ca2+/calmodulin-dependent protein kinase II in smooth muscle.

Z L Zhou 1, M Ikebe 1
PMCID: PMC1138298  PMID: 8172610

Abstract

Four novel isoforms of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) were found in rat aorta smooth muscle. Two of them were related to gamma-isoform of brain CaM kinase II (gamma-a). Differences in the primary structure of these isoforms were located in the variable region. One of them (gamma-b) contained 23 unique amino acid residues, whereas the other (gamma-c) did not contain this sequence. Both isoforms lacked the two segments (Val-316 to Gln-337 and Lys-353 to Leu-362) present in gamma-a. The DNA sequence of these gamma-isoforms except the variable region was exactly the same, suggesting that they are produced by alternative splicing. Another two isoforms were related to the delta-isoform of brain CaM kinase II (delta-a). delta-b contained a unique 11-residue sequence in the variable region whereas delta-c did not. As found for gamma-isoforms, the sequence analysis suggested that the three delta-isoforms are also produced by alternative splicing. Analysis of RNA by reverse transcription PCR confirmed the existence of specific messages for gamma-b, delta-a and delta-b. The variety of isoforms of CaM kinase II suggest that each isoform may play a specialized role in cell regulation.

Full text

PDF
490

Images in this article

493Figure 5
on p.493
493Figure 6
on p.493
493Figure 7
on p.493

Selected References

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

  1. Barnes W. M. Sequencing DNA with dideoxyribonucleotides as chain terminators: hints and strategies for big projects. Methods Enzymol. 1987;152:538–556. doi: 10.1016/0076-6879(87)52060-2. [DOI] [PubMed] [Google Scholar]
  2. Bennett M. K., Erondu N. E., Kennedy M. B. Purification and characterization of a calmodulin-dependent protein kinase that is highly concentrated in brain. J Biol Chem. 1983 Oct 25;258(20):12735–12744. [PubMed] [Google Scholar]
  3. Bennett M. K., Kennedy M. B. Deduced primary structure of the beta subunit of brain type II Ca2+/calmodulin-dependent protein kinase determined by molecular cloning. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1794–1798. doi: 10.1073/pnas.84.7.1794. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bulleit R. F., Bennett M. K., Molloy S. S., Hurley J. B., Kennedy M. B. Conserved and variable regions in the subunits of brain type II Ca2+/calmodulin-dependent protein kinase. Neuron. 1988 Mar;1(1):63–72. doi: 10.1016/0896-6273(88)90210-3. [DOI] [PubMed] [Google Scholar]
  5. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Fukunaga K., Yamamoto H., Matsui K., Higashi K., Miyamoto E. Purification and characterization of a Ca2+- and calmodulin-dependent protein kinase from rat brain. J Neurochem. 1982 Dec;39(6):1607–1617. doi: 10.1111/j.1471-4159.1982.tb07994.x. [DOI] [PubMed] [Google Scholar]
  8. Goldenring J. R., Gonzalez B., McGuire J. S., Jr, DeLorenzo R. J. Purification and characterization of a calmodulin-dependent kinase from rat brain cytosol able to phosphorylate tubulin and microtubule-associated proteins. J Biol Chem. 1983 Oct 25;258(20):12632–12640. [PubMed] [Google Scholar]
  9. Hanley R. M., Means A. R., Ono T., Kemp B. E., Burgin K. E., Waxham N., Kelly P. T. Functional analysis of a complementary DNA for the 50-kilodalton subunit of calmodulin kinase II. Science. 1987 Jul 17;237(4812):293–297. doi: 10.1126/science.3037704. [DOI] [PubMed] [Google Scholar]
  10. Hanson P. I., Schulman H. Neuronal Ca2+/calmodulin-dependent protein kinases. Annu Rev Biochem. 1992;61:559–601. doi: 10.1146/annurev.bi.61.070192.003015. [DOI] [PubMed] [Google Scholar]
  11. Hashimoto Y., Soderling T. R. Phosphorylation of smooth muscle myosin light chain kinase by Ca2+/calmodulin-dependent protein kinase II: comparative study of the phosphorylation sites. Arch Biochem Biophys. 1990 Apr;278(1):41–45. doi: 10.1016/0003-9861(90)90228-q. [DOI] [PubMed] [Google Scholar]
  12. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  13. Ikebe M., Reardon S. Phosphorylation of smooth muscle caldesmon by calmodulin-dependent protein kinase II. Identification of the phosphorylation sites. J Biol Chem. 1990 Oct 15;265(29):17607–17612. [PubMed] [Google Scholar]
  14. Ikebe M., Reardon S. Phosphorylation of smooth myosin light chain kinase by smooth muscle Ca2+/calmodulin-dependent multifunctional protein kinase. J Biol Chem. 1990 Jun 5;265(16):8975–8978. [PubMed] [Google Scholar]
  15. Ikebe M., Reardon S., Scott-Woo G. C., Zhou Z., Koda Y. Purification and characterization of calmodulin-dependent multifunctional protein kinase from smooth muscle: isolation of caldesmon kinase. Biochemistry. 1990 Dec 25;29(51):11242–11248. doi: 10.1021/bi00503a013. [DOI] [PubMed] [Google Scholar]
  16. Kanaseki T., Ikeuchi Y., Sugiura H., Yamauchi T. Structural features of Ca2+/calmodulin-dependent protein kinase II revealed by electron microscopy. J Cell Biol. 1991 Nov;115(4):1049–1060. doi: 10.1083/jcb.115.4.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kelly P. T., McGuinness T. L., Greengard P. Evidence that the major postsynaptic density protein is a component of a Ca2+/calmodulin-dependent protein kinase. Proc Natl Acad Sci U S A. 1984 Feb;81(3):945–949. doi: 10.1073/pnas.81.3.945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kennedy M. B., Bennett M. K., Erondu N. E. Biochemical and immunochemical evidence that the "major postsynaptic density protein" is a subunit of a calmodulin-dependent protein kinase. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7357–7361. doi: 10.1073/pnas.80.23.7357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kuret J., Schulman H. Purification and characterization of a Ca2+/calmodulin-dependent protein kinase from rat brain. Biochemistry. 1984 Nov 6;23(23):5495–5504. doi: 10.1021/bi00318a018. [DOI] [PubMed] [Google Scholar]
  20. Levine H., 3rd, Sahyoun N. E. Characterization of a soluble Mr-30,000 catalytic fragment of the neuronal calmodulin-dependent protein kinase II. Eur J Biochem. 1987 Nov 2;168(3):481–486. doi: 10.1111/j.1432-1033.1987.tb13442.x. [DOI] [PubMed] [Google Scholar]
  21. Lin C. R., Kapiloff M. S., Durgerian S., Tatemoto K., Russo A. F., Hanson P., Schulman H., Rosenfeld M. G. Molecular cloning of a brain-specific calcium/calmodulin-dependent protein kinase. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5962–5966. doi: 10.1073/pnas.84.16.5962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. McCarron J. G., McGeown J. G., Reardon S., Ikebe M., Fay F. S., Walsh J. V., Jr Calcium-dependent enhancement of calcium current in smooth muscle by calmodulin-dependent protein kinase II. Nature. 1992 May 7;357(6373):74–77. doi: 10.1038/357074a0. [DOI] [PubMed] [Google Scholar]
  23. Miller S. G., Kennedy M. B. Distinct forebrain and cerebellar isozymes of type II Ca2+/calmodulin-dependent protein kinase associate differently with the postsynaptic density fraction. J Biol Chem. 1985 Jul 25;260(15):9039–9046. [PubMed] [Google Scholar]
  24. Nghiem P., Saati S. M., Martens C. L., Gardner P., Schulman H. Cloning and analysis of two new isoforms of multifunctional Ca2+/calmodulin-dependent protein kinase. Expression in multiple human tissues. J Biol Chem. 1993 Mar 15;268(8):5471–5479. [PubMed] [Google Scholar]
  25. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stull J. T., Hsu L. C., Tansey M. G., Kamm K. E. Myosin light chain kinase phosphorylation in tracheal smooth muscle. J Biol Chem. 1990 Sep 25;265(27):16683–16690. [PubMed] [Google Scholar]
  27. Tobimatsu T., Fujisawa H. Tissue-specific expression of four types of rat calmodulin-dependent protein kinase II mRNAs. J Biol Chem. 1989 Oct 25;264(30):17907–17912. [PubMed] [Google Scholar]
  28. Tobimatsu T., Kameshita I., Fujisawa H. Molecular cloning of the cDNA encoding the third polypeptide (gamma) of brain calmodulin-dependent protein kinase II. J Biol Chem. 1988 Nov 5;263(31):16082–16086. [PubMed] [Google Scholar]
  29. Wang J., Best P. M. Inactivation of the sarcoplasmic reticulum calcium channel by protein kinase. Nature. 1992 Oct 22;359(6397):739–741. doi: 10.1038/359739a0. [DOI] [PubMed] [Google Scholar]
  30. Winder S. J., Walsh M. P. Smooth muscle calponin. Inhibition of actomyosin MgATPase and regulation by phosphorylation. J Biol Chem. 1990 Jun 15;265(17):10148–10155. [PubMed] [Google Scholar]
  31. Witcher D. R., Kovacs R. J., Schulman H., Cefali D. C., Jones L. R. Unique phosphorylation site on the cardiac ryanodine receptor regulates calcium channel activity. J Biol Chem. 1991 Jun 15;266(17):11144–11152. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES