Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1990 Jul 1;111(1):197–200. doi: 10.1083/jcb.111.1.197

Subcellular distribution of calcium-binding proteins and a calcium- ATPase in canine pancreas [published erratum appears in J Cell Biol 1990 Oct;111(4):1726]

PMCID: PMC2116152  PMID: 2142161

Abstract

Using a 45Ca blot-overlay assay, we monitored the subcellular fractionation pattern of several Ca binding proteins of apparent molecular masses 94, 61, and 59 kD. These proteins also appeared to stain blue with "Stains-All." Additionally, using a monoclonal antiserum raised against canine cardiac sarcoplasmic reticulum Ca- ATPase, we examined the subcellular distribution of a canine pancreatic 110-kD protein recognized by this antiserum. This protein had the same electrophoretic mobility as the cardiac protein against which the antiserum was raised. The three Ca binding proteins and the Ca-ATPase cofractionated into the rough microsomal fraction (RM), previously shown to consist of highly purified RER, in a pattern highly similar to that of the RER marker, ribophorin I. To provide further evidence for an RER localization, native RM were subjected to isopycnic flotation in sucrose gradients. The Ca binding proteins and the Ca-ATPase were found in dense fractions, along with ribophorin I. When RM were stripped of ribosomes with puromycin/high salt, the Ca binding proteins and the Ca- ATPase exhibited a shift to less dense fractions, as did ribophorin I. We conclude that, in pancreas, the Ca binding proteins and Ca-ATPase we detect are localized to the RER (conceivably a subcompartment of the RER) or, possibly, a structure intimately associated with the RER.

Full Text

The Full Text of this article is available as a PDF (622.7 KB).

Selected References

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

  1. Adelman M. R., Sabatini D. D., Blobel G. Ribosome-membrane interaction. Nondestructive disassembly of rat liver rough microsomes into ribosomal and membranous components. J Cell Biol. 1973 Jan;56(1):206–229. doi: 10.1083/jcb.56.1.206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bayerdörffer E., Streb H., Eckhardt L., Haase W., Schulz I. Characterization of calcium uptake into rough endoplasmic reticulum of rat pancreas. J Membr Biol. 1984;81(1):69–82. doi: 10.1007/BF01868811. [DOI] [PubMed] [Google Scholar]
  3. Berridge M. J., Irvine R. F. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. doi: 10.1038/341197a0. [DOI] [PubMed] [Google Scholar]
  4. Blobel G., Potter V. R. Studies on free and membrane-bound ribosomes in rat liver. I. Distribution as related to total cellular RNA. J Mol Biol. 1967 Jun 14;26(2):279–292. doi: 10.1016/0022-2836(67)90297-5. [DOI] [PubMed] [Google Scholar]
  5. Bolender R. P. Stereological analysis of the guinea pig pancreas. I. Analytical model and quantitative description of nonstimulated pancreatic exocrine cells. J Cell Biol. 1974 May;61(2):269–287. doi: 10.1083/jcb.61.2.269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Campbell K. P., MacLennan D. H., Jorgensen A. O. Staining of the Ca2+-binding proteins, calsequestrin, calmodulin, troponin C, and S-100, with the cationic carbocyanine dye "Stains-all". J Biol Chem. 1983 Sep 25;258(18):11267–11273. [PubMed] [Google Scholar]
  7. Damiani E., Spamer C., Heilmann C., Salvatori S., Margreth A. Endoplasmic reticulum of rat liver contains two proteins closely related to skeletal sarcoplasmic reticulum Ca-ATPase and calsequestrin. J Biol Chem. 1988 Jan 5;263(1):340–343. [PubMed] [Google Scholar]
  8. Ghosh T. K., Mullaney J. M., Tarazi F. I., Gill D. L. GTP-activated communication between distinct inositol 1,4,5-trisphosphate-sensitive and -insensitive calcium pools. Nature. 1989 Jul 20;340(6230):236–239. doi: 10.1038/340236a0. [DOI] [PubMed] [Google Scholar]
  9. Hashimoto S., Bruno B., Lew D. P., Pozzan T., Volpe P., Meldolesi J. Immunocytochemistry of calciosomes in liver and pancreas. J Cell Biol. 1988 Dec;107(6 Pt 2):2523–2531. doi: 10.1083/jcb.107.6.2523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jorgensen A. O., Arnold W., Pepper D. R., Kahl S. D., Mandel F., Campbell K. P. A monoclonal antibody to the Ca2+-ATPase of cardiac sarcoplasmic reticulum cross-reacts with slow type I but not with fast type II canine skeletal muscle fibers: an immunocytochemical and immunochemical study. Cell Motil Cytoskeleton. 1988;9(2):164–174. doi: 10.1002/cm.970090208. [DOI] [PubMed] [Google Scholar]
  11. Koch G., Smith M., Macer D., Webster P., Mortara R. Endoplasmic reticulum contains a common, abundant calcium-binding glycoprotein, endoplasmin. J Cell Sci. 1986 Dec;86:217–232. doi: 10.1242/jcs.86.1.217. [DOI] [PubMed] [Google Scholar]
  12. Kreibich G., Freienstein C. M., Pereyra B. N., Ulrich B. L., Sabatini D. D. Proteins of rough microsomal membranes related to ribosome binding. II. Cross-linking of bound ribosomes to specific membrane proteins exposed at the binding sites. J Cell Biol. 1978 May;77(2):488–506. doi: 10.1083/jcb.77.2.488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Maruyama K., Nonomura Y. High molecular weight calcium binding protein in the microsome of scallop striated muscle. J Biochem. 1984 Sep;96(3):859–870. doi: 10.1093/oxfordjournals.jbchem.a134904. [DOI] [PubMed] [Google Scholar]
  14. Mignery G. A., Südhof T. C., Takei K., De Camilli P. Putative receptor for inositol 1,4,5-trisphosphate similar to ryanodine receptor. Nature. 1989 Nov 9;342(6246):192–195. doi: 10.1038/342192a0. [DOI] [PubMed] [Google Scholar]
  15. Nigam S. K., Blobel G. Cyclic AMP-dependent protein kinase in canine pancreatic rough endoplasmic reticulum. J Biol Chem. 1989 Oct 5;264(28):16927–16932. [PubMed] [Google Scholar]
  16. Nigam S. K. Subcellular distribution of small GTP binding proteins in pancreas: identification of small GTP binding proteins in the rough endoplasmic reticulum. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1296–1299. doi: 10.1073/pnas.87.4.1296. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ross C. A., Meldolesi J., Milner T. A., Satoh T., Supattapone S., Snyder S. H. Inositol 1,4,5-trisphosphate receptor localized to endoplasmic reticulum in cerebellar Purkinje neurons. Nature. 1989 Jun 8;339(6224):468–470. doi: 10.1038/339468a0. [DOI] [PubMed] [Google Scholar]
  18. Rossier M. F., Capponi A. M., Vallotton M. B. The inositol 1,4,5-trisphosphate-binding site in adrenal cortical cells is distinct from the endoplasmic reticulum. J Biol Chem. 1989 Aug 25;264(24):14078–14084. [PubMed] [Google Scholar]
  19. Supattapone S., Danoff S. K., Theibert A., Joseph S. K., Steiner J., Snyder S. H. Cyclic AMP-dependent phosphorylation of a brain inositol trisphosphate receptor decreases its release of calcium. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8747–8750. doi: 10.1073/pnas.85.22.8747. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Van P. N., Peter F., Söling H. D. Four intracisternal calcium-binding glycoproteins from rat liver microsomes with high affinity for calcium. No indication for calsequestrin-like proteins in inositol 1,4,5-trisphosphate-sensitive calcium sequestering rat liver vesicles. J Biol Chem. 1989 Oct 15;264(29):17494–17501. [PubMed] [Google Scholar]
  21. Volpe P., Krause K. H., Hashimoto S., Zorzato F., Pozzan T., Meldolesi J., Lew D. P. "Calciosome," a cytoplasmic organelle: the inositol 1,4,5-trisphosphate-sensitive Ca2+ store of nonmuscle cells? Proc Natl Acad Sci U S A. 1988 Feb;85(4):1091–1095. doi: 10.1073/pnas.85.4.1091. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Waisman D. M., Salimath B. P., Anderson M. J. Isolation and characterization of CAB-63, a novel calcium-binding protein. J Biol Chem. 1985 Feb 10;260(3):1652–1660. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES