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. 1996 Oct;71(4):2130–2137. doi: 10.1016/S0006-3495(96)79413-4

Protons induce calsequestrin conformational changes.

C Hidalgo 1, P Donoso 1, P H Rodriguez 1
PMCID: PMC1233680  PMID: 8889188

Abstract

Calsequestrin, a high-capacity, intermediate-affinity, calcium-binding protein present in the lumen of sarcoplasmic reticulum, undergoes extensive calcium-induced conformational changes at neutral pH that cause distinct intrinsic fluorescence changes. The results reported in this work indicate that pH has a marked effect on these calcium-induced intrinsic fluorescence changes, as well as on calorimetric changes produced by the addition of Ca(2+) to calsequestrin. The addition of Ca(2+) at neutral pH produced a marked and cooperative increase in calsequestrin intrinsic fluorescence. In contrast, at pH 6.0 calsequestrin's intrinsic fluorescence was not affected by the addition of Ca(2+), and the same intrinsic fluorescence as that measured in millimolar calcium at neutral pH was obtained. The magnitude and the cooperativity of the calcium-induced intrinsic fluorescence changes decreased as either [H+] or [K+] increased. The evolution of heat production, determined by microcalorimetry, observed upon increasing the molar ratio of Ca(2+) to calsequestrin in 0.15 M KCl, decreased markedly as the pH decreased from pH 8.0 to pH 6.0, indicating that pH modifies the total heat content changes produced by Ca(2+). We propose that protons bind to calsequestrin and induce protein conformational changes that are responsible for the observed proton-induced intrinsic fluorescence and calorimetric changes.

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Selected References

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

  1. Aaron B. M., Oikawa K., Reithmeier R. A., Sykes B. D. Characterization of skeletal muscle calsequestrin by 1H NMR spectroscopy. J Biol Chem. 1984 Oct 10;259(19):11876–11881. [PubMed] [Google Scholar]
  2. Cala S. E., Jones L. R. Rapid purification of calsequestrin from cardiac and skeletal muscle sarcoplasmic reticulum vesicles by Ca2+-dependent elution from phenyl-sepharose. J Biol Chem. 1983 Oct 10;258(19):11932–11936. [PubMed] [Google Scholar]
  3. Cozens B., Reithmeier R. A. Size and shape of rabbit skeletal muscle calsequestrin. J Biol Chem. 1984 May 25;259(10):6248–6252. [PubMed] [Google Scholar]
  4. Damiani E., Margreth A. Specific protein-protein interactions of calsequestrin with junctional sarcoplasmic reticulum of skeletal muscle. Biochem Biophys Res Commun. 1990 Nov 15;172(3):1253–1259. doi: 10.1016/0006-291x(90)91584-f. [DOI] [PubMed] [Google Scholar]
  5. Damiani E., Salvatori S., Zorzato F., Margreth A. Characteristics of skeletal muscle calsequestrin: comparison of mammalian, amphibian and avian muscles. J Muscle Res Cell Motil. 1986 Oct;7(5):435–445. doi: 10.1007/BF01753586. [DOI] [PubMed] [Google Scholar]
  6. Donoso P., Prieto H., Hidalgo C. Luminal calcium regulates calcium release in triads isolated from frog and rabbit skeletal muscle. Biophys J. 1995 Feb;68(2):507–515. doi: 10.1016/S0006-3495(95)80212-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fliegel L., Ohnishi M., Carpenter M. R., Khanna V. K., Reithmeier R. A., MacLennan D. H. Amino acid sequence of rabbit fast-twitch skeletal muscle calsequestrin deduced from cDNA and peptide sequencing. Proc Natl Acad Sci U S A. 1987 Mar;84(5):1167–1171. doi: 10.1073/pnas.84.5.1167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Franzini-Armstrong C., Kenney L. J., Varriano-Marston E. The structure of calsequestrin in triads of vertebrate skeletal muscle: a deep-etch study. J Cell Biol. 1987 Jul;105(1):49–56. doi: 10.1083/jcb.105.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gilchrist J. S., Belcastro A. N., Katz S. Intraluminal Ca2+ dependence of Ca2+ and ryanodine-mediated regulation of skeletal muscle sarcoplasmic reticulum Ca2+ release. J Biol Chem. 1992 Oct 15;267(29):20850–20856. [PubMed] [Google Scholar]
  10. Guo W., Campbell K. P. Association of triadin with the ryanodine receptor and calsequestrin in the lumen of the sarcoplasmic reticulum. J Biol Chem. 1995 Apr 21;270(16):9027–9030. doi: 10.1074/jbc.270.16.9027. [DOI] [PubMed] [Google Scholar]
  11. Hartree E. F. Determination of protein: a modification of the Lowry method that gives a linear photometric response. Anal Biochem. 1972 Aug;48(2):422–427. doi: 10.1016/0003-2697(72)90094-2. [DOI] [PubMed] [Google Scholar]
  12. He Z., Dunker A. K., Wesson C. R., Trumble W. R. Ca(2+)-induced folding and aggregation of skeletal muscle sarcoplasmic reticulum calsequestrin. The involvement of the trifluoperazine-binding site. J Biol Chem. 1993 Nov 25;268(33):24635–24641. [PubMed] [Google Scholar]
  13. Hidalgo C., Donoso P. Luminal calcium regulation of calcium release from sarcoplasmic reticulum. Biosci Rep. 1995 Oct;15(5):387–397. doi: 10.1007/BF01788370. [DOI] [PubMed] [Google Scholar]
  14. Hidalgo C., Jorquera J., Tapia V., Donoso P. Triads and transverse tubules isolated from skeletal muscle contain high levels of inositol 1,4,5-trisphosphate. J Biol Chem. 1993 Jul 15;268(20):15111–15117. [PubMed] [Google Scholar]
  15. Ikemoto N., Antoniu B., Kang J. J., Mészáros L. G., Ronjat M. Intravesicular calcium transient during calcium release from sarcoplasmic reticulum. Biochemistry. 1991 May 28;30(21):5230–5237. doi: 10.1021/bi00235a017. [DOI] [PubMed] [Google Scholar]
  16. Ikemoto N., Bhatnagar G. M., Nagy B., Gergely J. Interaction of divalent cations with the 55,000-dalton protein component of the sarcoplasmic reticulum. Studies of fluorescence and circular dichroism. J Biol Chem. 1972 Dec 10;247(23):7835–7837. [PubMed] [Google Scholar]
  17. Ikemoto N., Nagy B., Bhatnagar G. M., Gergely J. Studies on a metal-binding protein of the sarcoplasmic reticulum. J Biol Chem. 1974 Apr 25;249(8):2357–2365. [PubMed] [Google Scholar]
  18. Ikemoto N., Ronjat M., Mészáros L. G., Koshita M. Postulated role of calsequestrin in the regulation of calcium release from sarcoplasmic reticulum. Biochemistry. 1989 Aug 8;28(16):6764–6771. doi: 10.1021/bi00442a033. [DOI] [PubMed] [Google Scholar]
  19. Jones L. R., Zhang L., Sanborn K., Jorgensen A. O., Kelley J. Purification, primary structure, and immunological characterization of the 26-kDa calsequestrin binding protein (junctin) from cardiac junctional sarcoplasmic reticulum. J Biol Chem. 1995 Dec 22;270(51):30787–30796. doi: 10.1074/jbc.270.51.30787. [DOI] [PubMed] [Google Scholar]
  20. Jorgensen A. O., Shen A. C., Campbell K. P. Ultrastructural localization of calsequestrin in adult rat atrial and ventricular muscle cells. J Cell Biol. 1985 Jul;101(1):257–268. doi: 10.1083/jcb.101.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kawasaki T., Kasai M. Regulation of calcium channel in sarcoplasmic reticulum by calsequestrin. Biochem Biophys Res Commun. 1994 Mar 30;199(3):1120–1127. doi: 10.1006/bbrc.1994.1347. [DOI] [PubMed] [Google Scholar]
  22. Krause K. H., Milos M., Luan-Rilliet Y., Lew D. P., Cox J. A. Thermodynamics of cation binding to rabbit skeletal muscle calsequestrin. Evidence for distinct Ca(2+)- and Mg(2+)-binding sites. J Biol Chem. 1991 May 25;266(15):9453–9459. [PubMed] [Google Scholar]
  23. MacLennan D. H., Wong P. T. Isolation of a calcium-sequestering protein from sarcoplasmic reticulum. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1231–1235. doi: 10.1073/pnas.68.6.1231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Maylie J., Irving M., Sizto N. L., Boyarsky G., Chandler W. K. Calcium signals recorded from cut frog twitch fibers containing tetramethylmurexide. J Gen Physiol. 1987 Jan;89(1):145–176. doi: 10.1085/jgp.89.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Meissner G. Isolation and characterization of two types of sarcoplasmic reticulum vesicles. Biochim Biophys Acta. 1975 Apr 21;389(1):51–68. doi: 10.1016/0005-2736(75)90385-5. [DOI] [PubMed] [Google Scholar]
  26. Meissner G., Young R. C. Proton permeability of sarcoplasmic reticulum vesicles. J Biol Chem. 1980 Jul 25;255(14):6814–6819. [PubMed] [Google Scholar]
  27. Mitchell R. D., Simmerman H. K., Jones L. R. Ca2+ binding effects on protein conformation and protein interactions of canine cardiac calsequestrin. J Biol Chem. 1988 Jan 25;263(3):1376–1381. [PubMed] [Google Scholar]
  28. Ohnishi M., Reithmeier R. A. Fragmentation of rabbit skeletal muscle calsequestrin: spectral and ion binding properties of the carboxyl-terminal region. Biochemistry. 1987 Nov 17;26(23):7458–7465. doi: 10.1021/bi00397a039. [DOI] [PubMed] [Google Scholar]
  29. Ostwald T. J., MacLennan D. H., Dorrington K. J. Effects of cation binding on the conformation of calsequestrin and the high affinity calcium-binding protein of sarcoplasmic reticulum. J Biol Chem. 1974 Sep 25;249(18):5867–5871. [PubMed] [Google Scholar]
  30. Pape P. C., Konishi M., Hollingworth S., Baylor S. M. Perturbation of sarcoplasmic reticulum calcium release and phenol red absorbance transients by large concentrations of fura-2 injected into frog skeletal muscle fibers. J Gen Physiol. 1990 Sep;96(3):493–516. doi: 10.1085/jgp.96.3.493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Saito A., Seiler S., Chu A., Fleischer S. Preparation and morphology of sarcoplasmic reticulum terminal cisternae from rabbit skeletal muscle. J Cell Biol. 1984 Sep;99(3):875–885. doi: 10.1083/jcb.99.3.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Slupsky J. R., Ohnishi M., Carpenter M. R., Reithmeier R. A. Characterization of cardiac calsequestrin. Biochemistry. 1987 Oct 6;26(20):6539–6544. doi: 10.1021/bi00394a038. [DOI] [PubMed] [Google Scholar]
  33. Somlyo A. V., Gonzalez-Serratos H. G., Shuman H., McClellan G., Somlyo A. P. Calcium release and ionic changes in the sarcoplasmic reticulum of tetanized muscle: an electron-probe study. J Cell Biol. 1981 Sep;90(3):577–594. doi: 10.1083/jcb.90.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Williams R. W., Beeler T. J. Secondary structure of calsequestrin in solutions and in crystals as determined by Raman spectroscopy. J Biol Chem. 1986 Sep 15;261(26):12408–12413. [PubMed] [Google Scholar]
  35. Wiseman T., Williston S., Brandts J. F., Lin L. N. Rapid measurement of binding constants and heats of binding using a new titration calorimeter. Anal Biochem. 1989 May 15;179(1):131–137. doi: 10.1016/0003-2697(89)90213-3. [DOI] [PubMed] [Google Scholar]
  36. Yano K., Zarain-Herzberg A. Sarcoplasmic reticulum calsequestrins: structural and functional properties. Mol Cell Biochem. 1994 Jun 15;135(1):61–70. doi: 10.1007/BF00925961. [DOI] [PubMed] [Google Scholar]

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