Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Jan;78(1):308–312. doi: 10.1073/pnas.78.1.308

Interactions of calmodulin with coated vesicles from brain.

C D Linden, J R Dedman, J G Chafouleas, A R Means, T F Roth
PMCID: PMC319042  PMID: 6941248

Abstract

Coated vesicles purified in the presence of calcium are enriched approximately 7-fold in calmodulin content relative to standard preparations isolated in the absence of free calcium. Radioiodinated calmodulin binds specifically to coated vesicles in vitro. Binding is saturable (Kd, 10 nM) and calcium dependent. Half-maximal binding occurs at 2.4 microM free Ca2+ whereas up to 1.2 mM Mg2+ has no effect on binding. Troponin C, a protein homologous to calmodulin, competes with binding of 125I-labeled calmodulin with 1/30th the affinity of native calmodulin. Chromatography of 2 M urea-solubilized coated vesicles on a calmodulin-Sepharose column demonstrated a Ca2+-dependent interaction of coated vesicle proteins and calmodulin. The properties of calmodulin binding to coated vesicles are comparable to those of calmodulin activities in other systems.

Full text

PDF
308

Images in this article

311Image
on p.311

Selected References

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

  1. Amphlett G. W., Vanaman T. C., Perry S. V. Effect of the troponin C-like protein from bovine brain (brain modulator protein) on the Mg2+-stimulated ATPase of skeletal muscle actinomyosin. FEBS Lett. 1976 Dec 15;72(1):163–168. doi: 10.1016/0014-5793(76)80836-8. [DOI] [PubMed] [Google Scholar]
  2. Blitz A. L., Fine R. E., Toselli P. A. Evidence that coated vesicles isolated from brain are calcium-sequestering organelles resembling sarcoplasmic reticulum. J Cell Biol. 1977 Oct;75(1):135–147. doi: 10.1083/jcb.75.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Chafouleas J. G., Dedman J. R., Munjaal R. P., Means A. R. Calmodulin. Development and application of a sensitive radioimmunoassay. J Biol Chem. 1979 Oct 25;254(20):10262–10267. [PubMed] [Google Scholar]
  6. DeLorenzo R. J., Freedman S. D., Yohe W. B., Maurer S. C. Stimulation of Ca2+-dependent neurotransmitter release and presynaptic nerve terminal protein phosphorylation by calmodulin and a calmodulin-like protein isolated from synaptic vesicles. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1838–1842. doi: 10.1073/pnas.76.4.1838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dedman J. R., Jackson R. L., Schreiber W. E., Means A. R. Sequence homology of the Ca2+-dependent regulator of cyclic nucleotide phosphodiesterase from rat testis with other Ca2+-binding proteins. J Biol Chem. 1978 Jan 25;253(2):343–346. [PubMed] [Google Scholar]
  8. Dedman J. R., Potter J. D., Jackson R. L., Johnson J. D., Means A. R. Physicochemical properties of rat testis Ca2+-dependent regulator protein of cyclic nucleotide phosphodiesterase. Relationship of Ca2+-binding, conformational changes, and phosphodiesterase activity. J Biol Chem. 1977 Dec 10;252(23):8415–8422. [PubMed] [Google Scholar]
  9. Dedman J. R., Potter J. D., Means A. R. Biological cross-reactivity of rat testis phosphodiesterase activator protein and rabbit skeletal muscle troponin-C. J Biol Chem. 1977 Apr 10;252(7):2437–2440. [PubMed] [Google Scholar]
  10. Dedman J. R., Welsh M. J., Means A. R. Ca2+-dependent regulator. Production and characterization of a monospecific antibody. J Biol Chem. 1978 Oct 25;253(20):7515–7521. [PubMed] [Google Scholar]
  11. Goldstein J. L., Anderson R. G., Brown M. S. Coated pits, coated vesicles, and receptor-mediated endocytosis. Nature. 1979 Jun 21;279(5715):679–685. doi: 10.1038/279679a0. [DOI] [PubMed] [Google Scholar]
  12. Gopinath R. M., Vincenzi F. F. Phosphodiesterase protein activator mimics red blood cell cytoplasmic activator of (Ca2+-Mg2+)ATPase. Biochem Biophys Res Commun. 1977 Aug 22;77(4):1203–1209. doi: 10.1016/s0006-291x(77)80107-1. [DOI] [PubMed] [Google Scholar]
  13. Grab D. J., Berzins K., Cohen R. S., Siekevitz P. Presence of calmodulin in postsynaptic densities isolated from canine cerebral cortex. J Biol Chem. 1979 Sep 10;254(17):8690–8696. [PubMed] [Google Scholar]
  14. Jarrett H. W., Kyte J. Human erythrocyte calmodulin. Further chemical characterization and the site of its interaction with the membrane. J Biol Chem. 1979 Sep 10;254(17):8237–8244. [PubMed] [Google Scholar]
  15. Jones H. P., Matthews J. C., Cormier M. J. Isolation and characterization of Ca2+-dependent modulator protein from the marine invertebrate Renilla reniformis. Biochemistry. 1979 Jan 9;18(1):55–60. doi: 10.1021/bi00568a009. [DOI] [PubMed] [Google Scholar]
  16. Kakiuchi S., Yamazaki R., Teshima Y., Uenishi K., Miyamoto E. Multiple cyclic nucleotide phosphodiesterase activities from rat tissues and occurrence of a calcium-plus-magnesium-ion-dependent phosphodiesterase and its protein activator. Biochem J. 1975 Jan;146(1):109–120. doi: 10.1042/bj1460109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kanaseki T., Kadota K. The "vesicle in a basket". A morphological study of the coated vesicle isolated from the nerve endings of the guinea pig brain, with special reference to the mechanism of membrane movements. J Cell Biol. 1969 Jul;42(1):202–220. doi: 10.1083/jcb.42.1.202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Keen J. H., Willingham M. C., Pastan I. H. Clathrin-coated vesicles: isolation, dissociation and factor-dependent reassociation of clathrin baskets. Cell. 1979 Feb;16(2):303–312. doi: 10.1016/0092-8674(79)90007-2. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Larsen F. L., Vincenzi F. F. Calcium transport across the plasma membrane: stimulation by calmodulin. Science. 1979 Apr 20;204(4390):306–309. doi: 10.1126/science.155309. [DOI] [PubMed] [Google Scholar]
  21. Marcum J. M., Dedman J. R., Brinkley B. R., Means A. R. Control of microtubule assembly-disassembly by calcium-dependent regulator protein. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3771–3775. doi: 10.1073/pnas.75.8.3771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pearse B. M. Clathrin: a unique protein associated with intracellular transfer of membrane by coated vesicles. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1255–1259. doi: 10.1073/pnas.73.4.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pearse B. M. Coated vesicles from pig brain: purification and biochemical characterization. J Mol Biol. 1975 Sep 5;97(1):93–98. doi: 10.1016/s0022-2836(75)80024-6. [DOI] [PubMed] [Google Scholar]
  24. Potter J. D., Gergely J. Troponin, tropomyosin, and actin interactions in the Ca2+ regulation of muscle contraction. Biochemistry. 1974 Jun 18;13(13):2697–2703. doi: 10.1021/bi00710a007. [DOI] [PubMed] [Google Scholar]
  25. ROTH T. F., PORTER K. R. YOLK PROTEIN UPTAKE IN THE OOCYTE OF THE MOSQUITO AEDES AEGYPTI. L. J Cell Biol. 1964 Feb;20:313–332. doi: 10.1083/jcb.20.2.313. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schook W., Puszkin S., Bloom W., Ores C., Kochwa S. Mechanochemical properties of brain clathrin: interactions with actin and alpha-actinin and polymerization into basketlike structures or filaments. Proc Natl Acad Sci U S A. 1979 Jan;76(1):116–120. doi: 10.1073/pnas.76.1.116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Schulman H., Greengard P. Ca2+-dependent protein phosphorylation system in membranes from various tissues, and its activation by "calcium-dependent regulator". Proc Natl Acad Sci U S A. 1978 Nov;75(11):5432–5436. doi: 10.1073/pnas.75.11.5432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Smoake J. A., Song S. Y., Cheung W. Y. Cyclic 3',5'-nucleotide phosphodiesterase. Distribution and developmental changes of the enzyme and its protein activator in mammalian tissues and cells. Biochim Biophys Acta. 1974 Apr 25;341(2):402–411. doi: 10.1016/0005-2744(74)90233-2. [DOI] [PubMed] [Google Scholar]
  29. Sobue K., Ichida S., Yoshida H., Yamazaki R., Kakiuchi S. Occurrence of a Ca2+- and modulator protein-activatable ATPase in the synaptic plasma membranes of brain. FEBS Lett. 1979 Mar 1;99(1):199–202. doi: 10.1016/0014-5793(79)80278-1. [DOI] [PubMed] [Google Scholar]
  30. Stevens F. C., Walsh M., Ho H. C., Teo T. S., Wang J. H. Comparison of calcium-binding proteins. Bovine heart and brain protein activators of cyclic nucleotide phosphodiesterase and rabbit skeletal muscle troponin C. J Biol Chem. 1976 Aug 10;251(15):4495–4500. [PubMed] [Google Scholar]
  31. Vandermeers A., Robberecht P., Vandermeers-Piret M. C., Rathé J., Christophe J. Specific binding of the calcium-dependent regulation protein to brain membranes from the guinea pig. Biochem Biophys Res Commun. 1978 Oct 30;84(4):1076–1081. doi: 10.1016/0006-291x(78)91693-5. [DOI] [PubMed] [Google Scholar]
  32. Waisman D., Stevens F. C., Wang J. H. The distribution of the Ca++-dependent protein activator of cyclic nucleotide phosphodiesterase in invertebrates. Biochem Biophys Res Commun. 1975 Aug 4;65(3):975–982. doi: 10.1016/s0006-291x(75)80481-5. [DOI] [PubMed] [Google Scholar]
  33. Watterson D. M., Harrelson W. G., Jr, Keller P. M., Sharief F., Vanaman T. C. Structural similarities between the Ca2+-dependent regulatory proteins of 3':5'-cyclic nucleotide phosphodiesterase and actomyosin ATPase. J Biol Chem. 1976 Aug 10;251(15):4501–4513. [PubMed] [Google Scholar]
  34. Woods J. W., Woodward M. P., Roth T. F. Common features of coated vesicles from dissimilar tissues: composition and structure. J Cell Sci. 1978 Apr;30:87–97. doi: 10.1242/jcs.30.1.87. [DOI] [PubMed] [Google Scholar]
  35. Woodward M. P., Roth T. F. Coated vesicles: characterization, selective dissociation, and reassembly. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4394–4398. doi: 10.1073/pnas.75.9.4394. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Woodward M. P., Roth T. F. Influence of buffer ions and divalent cations on coated vesicle disassembly and reassembly. J Supramol Struct. 1979;11(2):237–250. doi: 10.1002/jss.400110213. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES