Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1989 Feb;89(2):501–505. doi: 10.1104/pp.89.2.501

Purification and Characterization of Calmodulins from Papaver somniferum and Euphorbia lathyris

Marvin P Thompson 1, George J Piazza 1, Dorothy P Brower 1, Harold M Farrell Jr 1
PMCID: PMC1055871  PMID: 16666573

Abstract

Calmodulins (CaM) were isolated and characterized from two well-known latex producing plants, Papaver somniferum and Euphorbia lathyris. The molecular weights of both were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be 17,000 comparable to that of bovine brain CaM. Amino acid compositions also compared similarly with those of known CaMs, with regard to the presence of trimethyllysine and the ratio of phenylalanine to tyrosine. The Cornish-Bowden equation (SΔn) revealed strong statistical correlations of P. somniferum and E. lathyris CaM with those of other plants and animals, although their amino acid compositions were not identical. Both plant CaM stimulated CaM dependent cAMP phosphodiesterase: for Papaver somniferum the Ka was found to be 1.09 nanomolar and for Euphorbia lathyris, 2.01 nanomolar.

Full text

PDF
501

Images in this article

Selected References

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

  1. Anderson J. M., Charbonneau H., Jones H. P., McCann R. O., Cormier M. J. Characterization of the plant nicotinamide adenine dinucleotide kinase activator protein and its identification as calmodulin. Biochemistry. 1980 Jun 24;19(13):3113–3120. doi: 10.1021/bi00554a043. [DOI] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Butcher R. W. Cyclic 3',5'-nucleotide phosphodiesterase from bovine heart. Methods Enzymol. 1974;38:218–223. doi: 10.1016/0076-6879(74)38035-4. [DOI] [PubMed] [Google Scholar]
  4. Cheung W. Y. Cyclic 3',5'-nucleotide phosphodiesterase: pronounced stimulation by snake venom. Biochem Biophys Res Commun. 1967 Nov 30;29(4):478–482. doi: 10.1016/0006-291x(67)90508-6. [DOI] [PubMed] [Google Scholar]
  5. Cornish-Bowden A. Relating proteins by amino acid composition. Methods Enzymol. 1983;91:60–75. doi: 10.1016/s0076-6879(83)91011-x. [DOI] [PubMed] [Google Scholar]
  6. Eldik L. J., Grossman A. R., Iverson D. B., Watterson D. M. Isolation and characterization of calmodulin from spinach leaves and in vitro translation mixtures. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1912–1916. doi: 10.1073/pnas.77.4.1912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gopalakrishna R., Anderson W. B. Ca2+-induced hydrophobic site on calmodulin: application for purification of calmodulin by phenyl-Sepharose affinity chromatography. Biochem Biophys Res Commun. 1982 Jan 29;104(2):830–836. doi: 10.1016/0006-291x(82)90712-4. [DOI] [PubMed] [Google Scholar]
  8. Harmon A. C., Jarrett H. W., Cormier M. J. An enzymatic assay for calmodulins based on plant NAD kinase activity. Anal Biochem. 1984 Aug 15;141(1):168–178. doi: 10.1016/0003-2697(84)90441-x. [DOI] [PubMed] [Google Scholar]
  9. Ho C., Teo T. S., Desai R., Wang J. H. Catalytic and regulatory properties of two forms of bovine heart cyclic nucleotide phosphodiesterase. Biochim Biophys Acta. 1976 Apr 8;429(2):461–473. doi: 10.1016/0005-2744(76)90294-1. [DOI] [PubMed] [Google Scholar]
  10. Jarrett H. W., Charbonneau H., Anderson J. M., McCann R. O., Cormier M. J. Plant calmodulin and the regulation of NAD kinase. Ann N Y Acad Sci. 1980;356:119–129. doi: 10.1111/j.1749-6632.1980.tb29605.x. [DOI] [PubMed] [Google Scholar]
  11. Kretsinger R. H. Crystallographic studies of calmodulin and homologs. Ann N Y Acad Sci. 1980;356:14–19. doi: 10.1111/j.1749-6632.1980.tb29594.x. [DOI] [PubMed] [Google Scholar]
  12. Lukas T. J., Iverson D. B., Schleicher M., Watterson D. M. Structural characterization of a higher plant calmodulin : spinacia oleracea. Plant Physiol. 1984 Jul;75(3):788–795. doi: 10.1104/pp.75.3.788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Muto S., Miyachi S. Properties of a Protein Activator of NAD Kinase from Plants. Plant Physiol. 1977 Jan;59(1):55–60. doi: 10.1104/pp.59.1.55. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nessler C. L., Allen R. D., Galewsky S. Identification and characterization of latex-specific proteins in opium poppy. Plant Physiol. 1985 Oct;79(2):499–504. doi: 10.1104/pp.79.2.499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Piazza G. J., Saggese E. J., Spletzer K. M. Triterpene Biosynthesis in the Latex of Euphorbia lathyris: Effect of Calmodulin Antagonists and Chlorinated Phenoxy Compounds. Plant Physiol. 1987 Jan;83(1):177–180. doi: 10.1104/pp.83.1.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schleicher M., Lukas T. J., Watterson D. M. Further Characterization of Calmodulin from the Monocotyledon Barley (Hordeum vulgare). Plant Physiol. 1983 Nov;73(3):666–670. doi: 10.1104/pp.73.3.666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schreiber W. E., Sasagawa T., Titani K., Wade R. D., Malencik D., Fischer E. H. Human brain calmodulin: isolation, characterization, and sequence of a half-molecule fragment. Biochemistry. 1981 Sep 1;20(18):5239–5245. doi: 10.1021/bi00521a022. [DOI] [PubMed] [Google Scholar]
  18. Thompson M. P., Brower D. P., Farrell H. M., Jr Absence of detectable calmodulin in cows' milk by a modified gel electrophoresis method. J Dairy Sci. 1987 Jun;70(6):1134–1139. doi: 10.3168/jds.S0022-0302(87)80123-6. [DOI] [PubMed] [Google Scholar]
  19. Thompson M. P., Piazza G. J., Brower D. P., Bingham E. W., Farrell H. M., Jr Isolation and characterization of bovine mammary calmodulin. J Dairy Sci. 1987 Aug;70(8):1551–1556. doi: 10.3168/jds.S0022-0302(87)80182-0. [DOI] [PubMed] [Google Scholar]
  20. Toda H., Yazawa M., Sakiyama F., Yagi K. Amino acid sequence of calmodulin from wheat germ. J Biochem. 1985 Sep;98(3):833–842. doi: 10.1093/oxfordjournals.jbchem.a135342. [DOI] [PubMed] [Google Scholar]
  21. Wang J. H., Waisman D. M. Calmodulin and its role in the second-messenger system. Curr Top Cell Regul. 1979;15:47–107. doi: 10.1016/b978-0-12-152815-7.50006-5. [DOI] [PubMed] [Google Scholar]
  22. Watterson D. M., Iverson D. B., Van Eldik L. J. Spinach calmodulin: isolation, characterization, and comparison with vertebrate calmodulins. Biochemistry. 1980 Dec 9;19(25):5762–5768. doi: 10.1021/bi00566a015. [DOI] [PubMed] [Google Scholar]
  23. Watterson D. M., Sharief F., Vanaman T. C. The complete amino acid sequence of the Ca2+-dependent modulator protein (calmodulin) of bovine brain. J Biol Chem. 1980 Feb 10;255(3):962–975. [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES