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. 1984 Dec 1;3(12):2969–2974. doi: 10.1002/j.1460-2075.1984.tb02242.x

Cellular distribution of three mammalian Ca2+-binding proteins related to Torpedo calelectrin.

M Geisow, J Childs, B Dash, A Harris, G Panayotou, T Südhof, J H Walker
PMCID: PMC557799  PMID: 6241147

Abstract

Addition of Ca2+ to post-microsomal fractions of bovine adrenal or liver produced a sedimentable complex of membrane vesicles and cytoplasmic proteins. Proteins with apparent mol. wts. 70 000, 36 000 and 32 500 were solubilized from this complex by Ca2+ chelation. The 36 000 mol. wt. protein (p36) was immunoprecipitated by an antiserum specific for pp36, a major substrate for Rous sarcoma virus src-gene tyrosine kinase. This protein was present in many mesenchymal cells and associated with membrane cytoskeleton of bovine fibroblasts in a Ca2+-dependent manner. The 70 000 and 32 500 mol. wt. proteins were widely distributed in established cell lines, but were not clearly associated with cell organelles in tissue sections, nor retained in cytoskeleton preparations. On immunoblots p36 reacted strongly with antibodies produced against the electric fish protein Torpedo calelectrin and the similar Ca2+-binding properties and subunit mol. wts. of these proteins suggests that they might be functionally related. Since Torpedo calelectrin, p70, p36 and p32.5 were bound by lipid vesicles or microsomal membranes at micromolar free Ca2+ concentrations, regulated association with intrinsic membrane components may be involved in the functions of these widespread proteins.

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Selected References

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

  1. Burgoyne R. D., Geisow M. J. Specific binding of 125I-calmodulin to and protein phosphorylation in adrenal chromaffin granule membranes. FEBS Lett. 1981 Aug 17;131(1):127–131. doi: 10.1016/0014-5793(81)80903-9. [DOI] [PubMed] [Google Scholar]
  2. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  3. Courtneidge S., Ralston R., Alitalo K., Bishop J. M. Subcellular location of an abundant substrate (p36) for tyrosine-specific protein kinases. Mol Cell Biol. 1983 Mar;3(3):340–350. doi: 10.1128/mcb.3.3.340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Creutz C. E., Dowling L. G., Sando J. J., Villar-Palasi C., Whipple J. H., Zaks W. J. Characterization of the chromobindins. Soluble proteins that bind to the chromaffin granule membrane in the presence of Ca2+. J Biol Chem. 1983 Dec 10;258(23):14664–14674. [PubMed] [Google Scholar]
  5. Erikson E., Tomasiewicz H. G., Erikson R. L. Biochemical characterization of a 34-kilodalton normal cellular substrate of pp60v-src and an associated 6-kilodalton protein. Mol Cell Biol. 1984 Jan;4(1):77–85. doi: 10.1128/mcb.4.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Geiger B. Membrane-cytoskeleton interaction. Biochim Biophys Acta. 1983 Aug 11;737(3-4):305–341. doi: 10.1016/0304-4157(83)90005-9. [DOI] [PubMed] [Google Scholar]
  7. Geisow M. J., Burgoyne R. D. Calcium-dependent binding of cytosolic proteins by chromaffin granules from adrenal medulla. J Neurochem. 1982 Jun;38(6):1735–1741. doi: 10.1111/j.1471-4159.1982.tb06656.x. [DOI] [PubMed] [Google Scholar]
  8. Gerke V., Weber K. Identity of p36K phosphorylated upon Rous sarcoma virus transformation with a protein purified from brush borders; calcium-dependent binding to non-erythroid spectrin and F-actin. EMBO J. 1984 Jan;3(1):227–233. doi: 10.1002/j.1460-2075.1984.tb01789.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greenberg M. E., Edelman G. M. Comparison of the 34,000-Da pp60src substrate and a 38,000-Da phosphoprotein identified by monoclonal antibodies. J Biol Chem. 1983 Jul 10;258(13):8497–8502. [PubMed] [Google Scholar]
  10. Hunter T., Cooper J. A. Epidermal growth factor induces rapid tyrosine phosphorylation of proteins in A431 human tumor cells. Cell. 1981 Jun;24(3):741–752. doi: 10.1016/0092-8674(81)90100-8. [DOI] [PubMed] [Google Scholar]
  11. Kikkawa U., Takai Y., Minakuchi R., Inohara S., Nishizuka Y. Calcium-activated, phospholipid-dependent protein kinase from rat brain. Subcellular distribution, purification, and properties. J Biol Chem. 1982 Nov 25;257(22):13341–13348. [PubMed] [Google Scholar]
  12. Kilimann M. W., Isenberg G. Actin filament capping protein from bovine brain. EMBO J. 1982;1(7):889–894. doi: 10.1002/j.1460-2075.1982.tb01265.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Moore P. B., Dedman J. R. Calcium-dependent protein binding to phenothiazine columns. J Biol Chem. 1982 Aug 25;257(16):9663–9667. [PubMed] [Google Scholar]
  14. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  15. Owens R. J., Gallagher C. J., Crumpton M. J. Cellular distribution of p68, a new calcium-binding protein from lymphocytes. EMBO J. 1984 May;3(5):945–952. doi: 10.1002/j.1460-2075.1984.tb01912.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Radke K., Martin G. S. Transformation by Rous sarcoma virus: effects of src gene expression on the synthesis and phosphorylation of cellular polypeptides. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5212–5216. doi: 10.1073/pnas.76.10.5212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Südhof T. C., Ebbecke M., Walker J. H., Fritsche U., Boustead C. Isolation of mammalian calelectrins: a new class of ubiquitous Ca2+-regulated proteins. Biochemistry. 1984 Mar 13;23(6):1103–1109. doi: 10.1021/bi00301a010. [DOI] [PubMed] [Google Scholar]
  18. Südhof T. C., Walker J. H., Obrocki J. Calelectrin self-aggregates and promotes membrane aggregation in the presence of calcium. EMBO J. 1982;1(10):1167–1170. doi: 10.1002/j.1460-2075.1982.tb00008.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Südhof T. C., Zimmermann C. W., Walker J. H. Calelectrin in human blood cells. Eur J Cell Biol. 1983 May;30(2):214–218. [PubMed] [Google Scholar]
  20. Walker J. H. Isolation from cholinergic synapses of a protein that binds to membranes in a calcium-dependent manner. J Neurochem. 1982 Sep;39(3):815–823. doi: 10.1111/j.1471-4159.1982.tb07965.x. [DOI] [PubMed] [Google Scholar]
  21. Walker J. H., Obrocki J., Südhof T. C. Calelectrin, a calcium-dependent membrane-binding protein associated with secretory granules in Torpedo cholinergic electromotor nerve endings and rat adrenal medulla. J Neurochem. 1983 Jul;41(1):139–145. doi: 10.1111/j.1471-4159.1983.tb11825.x. [DOI] [PubMed] [Google Scholar]
  22. Weingarten M. D., Lockwood A. H., Hwo S. Y., Kirschner M. W. A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A. 1975 May;72(5):1858–1862. doi: 10.1073/pnas.72.5.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wilson S. P., Viveros O. H. Primary culture of adrenal medullary chromaffin cells in a chemically defined medium. Exp Cell Res. 1981 May;133(1):159–169. doi: 10.1016/0014-4827(81)90366-9. [DOI] [PubMed] [Google Scholar]

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