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. 1999 Oct;77(4):1936–1944. doi: 10.1016/S0006-3495(99)77035-9

Structure of the skeletal muscle calcium release channel activated with Ca2+ and AMP-PCP.

I I Serysheva 1, M Schatz 1, M van Heel 1, W Chiu 1, S L Hamilton 1
PMCID: PMC1300475  PMID: 10512814

Abstract

The functional state of the skeletal muscle Ca2+ release channel is modulated by a number of endogenous molecules during excitation-contraction. Using electron cryomicroscopy and angular reconstitution techniques, we determined the three-dimensional (3D) structure of the skeletal muscle Ca2+ release channel activated by a nonhydrolyzable analog of ATP in the presence of Ca2+. These ligands together produce almost maximum activation of the channel and drive the channel population toward a predominately open state. The resulting 30-A 3D reconstruction reveals long-range conformational changes in the cytoplasmic region that might affect the interaction of the Ca2+ release channel with the t-tubule voltage sensor. In addition, a central opening and mass movements, detected in the transmembrane domain of both the Ca(2+)- and the Ca2+/nucleotide-activated channels, suggest a mechanism for channel opening similar to opening-closing of the iris in a camera diaphragm.

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Selected References

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  1. Block B. A., Imagawa T., Campbell K. P., Franzini-Armstrong C. Structural evidence for direct interaction between the molecular components of the transverse tubule/sarcoplasmic reticulum junction in skeletal muscle. J Cell Biol. 1988 Dec;107(6 Pt 2):2587–2600. doi: 10.1083/jcb.107.6.2587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Callaway C., Seryshev A., Wang J. P., Slavik K. J., Needleman D. H., Cantu C., 3rd, Wu Y., Jayaraman T., Marks A. R., Hamilton S. L. Localization of the high and low affinity [3H]ryanodine binding sites on the skeletal muscle Ca2+ release channel. J Biol Chem. 1994 Jun 3;269(22):15876–15884. [PubMed] [Google Scholar]
  3. Chu A., Díaz-Muñoz M., Hawkes M. J., Brush K., Hamilton S. L. Ryanodine as a probe for the functional state of the skeletal muscle sarcoplasmic reticulum calcium release channel. Mol Pharmacol. 1990 May;37(5):735–741. [PubMed] [Google Scholar]
  4. Copello J. A., Barg S., Onoue H., Fleischer S. Heterogeneity of Ca2+ gating of skeletal muscle and cardiac ryanodine receptors. Biophys J. 1997 Jul;73(1):141–156. doi: 10.1016/S0006-3495(97)78055-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Coronado R., Morrissette J., Sukhareva M., Vaughan D. M. Structure and function of ryanodine receptors. Am J Physiol. 1994 Jun;266(6 Pt 1):C1485–C1504. doi: 10.1152/ajpcell.1994.266.6.C1485. [DOI] [PubMed] [Google Scholar]
  6. Du G. G., MacLennan D. H. Functional consequences of mutations of conserved, polar amino acids in transmembrane sequences of the Ca2+ release channel (ryanodine receptor) of rabbit skeletal muscle sarcoplasmic reticulum. J Biol Chem. 1998 Nov 27;273(48):31867–31872. doi: 10.1074/jbc.273.48.31867. [DOI] [PubMed] [Google Scholar]
  7. Díaz-Muñoz M., Hamilton S. L., Kaetzel M. A., Hazarika P., Dedman J. R. Modulation of Ca2+ release channel activity from sarcoplasmic reticulum by annexin VI (67-kDa calcimedin). J Biol Chem. 1990 Sep 15;265(26):15894–15899. [PubMed] [Google Scholar]
  8. Escobar A. L., Monck J. R., Fernandez J. M., Vergara J. L. Localization of the site of Ca2+ release at the level of a single sarcomere in skeletal muscle fibres. Nature. 1994 Feb 24;367(6465):739–741. doi: 10.1038/367739a0. [DOI] [PubMed] [Google Scholar]
  9. Fleig A., Takeshima H., Penner R. Absence of Ca2+ current facilitation in skeletal muscle of transgenic mice lacking the type 1 ryanodine receptor. J Physiol. 1996 Oct 15;496(Pt 2):339–345. doi: 10.1113/jphysiol.1996.sp021689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fleischer S., Inui M. Biochemistry and biophysics of excitation-contraction coupling. Annu Rev Biophys Biophys Chem. 1989;18:333–364. doi: 10.1146/annurev.bb.18.060189.002001. [DOI] [PubMed] [Google Scholar]
  11. Hawkes M. J., Díaz-Muñoz M., Hamilton S. L. A procedure for purification of the ryanodine receptor from skeletal muscle. Membr Biochem. 1989;8(3):133–145. doi: 10.3109/09687688909025827. [DOI] [PubMed] [Google Scholar]
  12. Holmberg S. R., Williams A. J. The cardiac sarcoplasmic reticulum calcium-release channel: modulation of ryanodine binding and single-channel activity. Biochim Biophys Acta. 1990 Feb 28;1022(2):187–193. doi: 10.1016/0005-2736(90)90113-3. [DOI] [PubMed] [Google Scholar]
  13. Ikemoto N., Antoniu B., Mészáros L. G. Rapid flow chemical quench studies of calcium release from isolated sarcoplasmic reticulum. J Biol Chem. 1985 Nov 15;260(26):14096–14100. [PubMed] [Google Scholar]
  14. Jayaraman T., Brillantes A. M., Timerman A. P., Fleischer S., Erdjument-Bromage H., Tempst P., Marks A. R. FK506 binding protein associated with the calcium release channel (ryanodine receptor). J Biol Chem. 1992 May 15;267(14):9474–9477. [PubMed] [Google Scholar]
  15. Kang J. J., Tarcsafalvi A., Carlos A. D., Fujimoto E., Shahrokh Z., Thevenin B. J., Shohet S. B., Ikemoto N. Conformational changes in the foot protein of the sarcoplasmic reticulum assessed by site-directed fluorescent labeling. Biochemistry. 1992 Mar 31;31(12):3288–3293. doi: 10.1021/bi00127a034. [DOI] [PubMed] [Google Scholar]
  16. Lai F. A., Misra M., Xu L., Smith H. A., Meissner G. The ryanodine receptor-Ca2+ release channel complex of skeletal muscle sarcoplasmic reticulum. Evidence for a cooperatively coupled, negatively charged homotetramer. J Biol Chem. 1989 Oct 5;264(28):16776–16785. [PubMed] [Google Scholar]
  17. Leong P., MacLennan D. H. The cytoplasmic loops between domains II and III and domains III and IV in the skeletal muscle dihydropyridine receptor bind to a contiguous site in the skeletal muscle ryanodine receptor. J Biol Chem. 1998 Nov 6;273(45):29958–29964. doi: 10.1074/jbc.273.45.29958. [DOI] [PubMed] [Google Scholar]
  18. Ma J. Desensitization of the skeletal muscle ryanodine receptor: evidence for heterogeneity of calcium release channels. Biophys J. 1995 Mar;68(3):893–899. doi: 10.1016/S0006-3495(95)80265-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ma J., Fill M., Knudson C. M., Campbell K. P., Coronado R. Ryanodine receptor of skeletal muscle is a gap junction-type channel. Science. 1988 Oct 7;242(4875):99–102. doi: 10.1126/science.2459777. [DOI] [PubMed] [Google Scholar]
  20. McGrew S. G., Wolleben C., Siegl P., Inui M., Fleischer S. Positive cooperativity of ryanodine binding to the calcium release channel of sarcoplasmic reticulum from heart and skeletal muscle. Biochemistry. 1989 Feb 21;28(4):1686–1691. doi: 10.1021/bi00430a039. [DOI] [PubMed] [Google Scholar]
  21. Meissner G., Darling E., Eveleth J. Kinetics of rapid Ca2+ release by sarcoplasmic reticulum. Effects of Ca2+, Mg2+, and adenine nucleotides. Biochemistry. 1986 Jan 14;25(1):236–244. doi: 10.1021/bi00349a033. [DOI] [PubMed] [Google Scholar]
  22. Meissner G. Evidence of a role for calmodulin in the regulation of calcium release from skeletal muscle sarcoplasmic reticulum. Biochemistry. 1986 Jan 14;25(1):244–251. doi: 10.1021/bi00349a034. [DOI] [PubMed] [Google Scholar]
  23. Meissner G., Henderson J. S. Rapid calcium release from cardiac sarcoplasmic reticulum vesicles is dependent on Ca2+ and is modulated by Mg2+, adenine nucleotide, and calmodulin. J Biol Chem. 1987 Mar 5;262(7):3065–3073. [PubMed] [Google Scholar]
  24. Meissner G. Ryanodine activation and inhibition of the Ca2+ release channel of sarcoplasmic reticulum. J Biol Chem. 1986 May 15;261(14):6300–6306. [PubMed] [Google Scholar]
  25. Meissner G. Ryanodine receptor/Ca2+ release channels and their regulation by endogenous effectors. Annu Rev Physiol. 1994;56:485–508. doi: 10.1146/annurev.ph.56.030194.002413. [DOI] [PubMed] [Google Scholar]
  26. Meissner G., el-Hashem A. Ryanodine as a functional probe of the skeletal muscle sarcoplasmic reticulum Ca2+ release channel. Mol Cell Biochem. 1992 Sep 8;114(1-2):119–123. doi: 10.1007/BF00240306. [DOI] [PubMed] [Google Scholar]
  27. Nagasaki K., Fleischer S. Ryanodine sensitivity of the calcium release channel of sarcoplasmic reticulum. Cell Calcium. 1988 Feb;9(1):1–7. doi: 10.1016/0143-4160(88)90032-2. [DOI] [PubMed] [Google Scholar]
  28. Nakai J., Dirksen R. T., Nguyen H. T., Pessah I. N., Beam K. G., Allen P. D. Enhanced dihydropyridine receptor channel activity in the presence of ryanodine receptor. Nature. 1996 Mar 7;380(6569):72–75. doi: 10.1038/380072a0. [DOI] [PubMed] [Google Scholar]
  29. Ogawa Y. Role of ryanodine receptors. Crit Rev Biochem Mol Biol. 1994;29(4):229–274. doi: 10.3109/10409239409083482. [DOI] [PubMed] [Google Scholar]
  30. Ohkusa T., Kang J. J., Morii M., Ikemoto N. Conformational change of the foot protein of sarcoplasmic reticulum as an initial event of calcium release. J Biochem. 1991 Apr;109(4):609–615. doi: 10.1093/oxfordjournals.jbchem.a123428. [DOI] [PubMed] [Google Scholar]
  31. Orlova E. V., Dube P., Harris J. R., Beckman E., Zemlin F., Markl J., van Heel M. Structure of keyhole limpet hemocyanin type 1 (KLH1) at 15 A resolution by electron cryomicroscopy and angular reconstitution. J Mol Biol. 1997 Aug 22;271(3):417–437. doi: 10.1006/jmbi.1997.1182. [DOI] [PubMed] [Google Scholar]
  32. Orlova E. V., Serysheva I. I., van Heel M., Hamilton S. L., Chiu W. Two structural configurations of the skeletal muscle calcium release channel. Nat Struct Biol. 1996 Jun;3(6):547–552. doi: 10.1038/nsb0696-547. [DOI] [PubMed] [Google Scholar]
  33. Pessah I. N., Zimanyi I. Characterization of multiple [3H]ryanodine binding sites on the Ca2+ release channel of sarcoplasmic reticulum from skeletal and cardiac muscle: evidence for a sequential mechanism in ryanodine action. Mol Pharmacol. 1991 May;39(5):679–689. [PubMed] [Google Scholar]
  34. Radermacher M., Rao V., Grassucci R., Frank J., Timerman A. P., Fleischer S., Wagenknecht T. Cryo-electron microscopy and three-dimensional reconstruction of the calcium release channel/ryanodine receptor from skeletal muscle. J Cell Biol. 1994 Oct;127(2):411–423. doi: 10.1083/jcb.127.2.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rios E., Brum G. Involvement of dihydropyridine receptors in excitation-contraction coupling in skeletal muscle. Nature. 1987 Feb 19;325(6106):717–720. doi: 10.1038/325717a0. [DOI] [PubMed] [Google Scholar]
  36. Rousseau E., Pinkos J., Savaria D. Functional sensitivity of the native skeletal Ca(2+)-release channel to divalent cations and the Mg-ATP complex. Can J Physiol Pharmacol. 1992 Mar;70(3):394–402. doi: 10.1139/y92-049. [DOI] [PubMed] [Google Scholar]
  37. Rousseau E., Smith J. S., Meissner G. Ryanodine modifies conductance and gating behavior of single Ca2+ release channel. Am J Physiol. 1987 Sep;253(3 Pt 1):C364–C368. doi: 10.1152/ajpcell.1987.253.3.C364. [DOI] [PubMed] [Google Scholar]
  38. Sass H. J., Büldt G., Beckmann E., Zemlin F., van Heel M., Zeitler E., Rosenbusch J. P., Dorset D. L., Massalski A. Densely packed beta-structure at the protein-lipid interface of porin is revealed by high-resolution cryo-electron microscopy. J Mol Biol. 1989 Sep 5;209(1):171–175. doi: 10.1016/0022-2836(89)90180-0. [DOI] [PubMed] [Google Scholar]
  39. Schatz M., Orlova E. V., Dube P., Jäger J., van Heel M. Structure of Lumbricus terrestris hemoglobin at 30 A resolution determined using angular reconstitution. J Struct Biol. 1995 Jan-Feb;114(1):28–40. doi: 10.1006/jsbi.1995.1003. [DOI] [PubMed] [Google Scholar]
  40. Serysheva I. I., Orlova E. V., Chiu W., Sherman M. B., Hamilton S. L., van Heel M. Electron cryomicroscopy and angular reconstitution used to visualize the skeletal muscle calcium release channel. Nat Struct Biol. 1995 Jan;2(1):18–24. doi: 10.1038/nsb0195-18. [DOI] [PubMed] [Google Scholar]
  41. Smith J. S., Coronado R., Meissner G. Single channel measurements of the calcium release channel from skeletal muscle sarcoplasmic reticulum. Activation by Ca2+ and ATP and modulation by Mg2+. J Gen Physiol. 1986 Nov;88(5):573–588. doi: 10.1085/jgp.88.5.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Smith J. S., Imagawa T., Ma J., Fill M., Campbell K. P., Coronado R. Purified ryanodine receptor from rabbit skeletal muscle is the calcium-release channel of sarcoplasmic reticulum. J Gen Physiol. 1988 Jul;92(1):1–26. doi: 10.1085/jgp.92.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Stern M. D., Pizarro G., Ríos E. Local control model of excitation-contraction coupling in skeletal muscle. J Gen Physiol. 1997 Oct;110(4):415–440. doi: 10.1085/jgp.110.4.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Takeshima H., Nishimura S., Matsumoto T., Ishida H., Kangawa K., Minamino N., Matsuo H., Ueda M., Hanaoka M., Hirose T. Primary structure and expression from complementary DNA of skeletal muscle ryanodine receptor. Nature. 1989 Jun 8;339(6224):439–445. doi: 10.1038/339439a0. [DOI] [PubMed] [Google Scholar]
  45. Timerman A. P., Ogunbumni E., Freund E., Wiederrecht G., Marks A. R., Fleischer S. The calcium release channel of sarcoplasmic reticulum is modulated by FK-506-binding protein. Dissociation and reconstitution of FKBP-12 to the calcium release channel of skeletal muscle sarcoplasmic reticulum. J Biol Chem. 1993 Nov 5;268(31):22992–22999. [PubMed] [Google Scholar]
  46. Timerman A. P., Wiederrecht G., Marcy A., Fleischer S. Characterization of an exchange reaction between soluble FKBP-12 and the FKBP.ryanodine receptor complex. Modulation by FKBP mutants deficient in peptidyl-prolyl isomerase activity. J Biol Chem. 1995 Feb 10;270(6):2451–2459. doi: 10.1074/jbc.270.6.2451. [DOI] [PubMed] [Google Scholar]
  47. Tinker A., Sutko J. L., Ruest L., Deslongchamps P., Welch W., Airey J. A., Gerzon K., Bidasee K. R., Besch H. R., Jr, Williams A. J. Electrophysiological effects of ryanodine derivatives on the sheep cardiac sarcoplasmic reticulum calcium-release channel. Biophys J. 1996 May;70(5):2110–2119. doi: 10.1016/S0006-3495(96)79777-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Wang J. P., Needleman D. H., Hamilton S. L. Relationship of low affinity [3]ryanodine binding sites to high affinity sites on the skeletal muscle Ca2+ release channel. J Biol Chem. 1993 Oct 5;268(28):20974–20982. [PubMed] [Google Scholar]
  49. Witcher D. R., McPherson P. S., Kahl S. D., Lewis T., Bentley P., Mullinnix M. J., Windass J. D., Campbell K. P. Photoaffinity labeling of the ryanodine receptor/Ca2+ release channel with an azido derivative of ryanodine. J Biol Chem. 1994 May 6;269(18):13076–13079. [PubMed] [Google Scholar]
  50. Zhou Z. H., Hardt S., Wang B., Sherman M. B., Jakana J., Chiu W. CTF determination of images of ice-embedded single particles using a graphics interface. J Struct Biol. 1996 Jan-Feb;116(1):216–222. doi: 10.1006/jsbi.1996.0033. [DOI] [PubMed] [Google Scholar]
  51. Zhou Z. H., Prasad B. V., Jakana J., Rixon F. J., Chiu W. Protein subunit structures in the herpes simplex virus A-capsid determined from 400 kV spot-scan electron cryomicroscopy. J Mol Biol. 1994 Sep 30;242(4):456–469. doi: 10.1006/jmbi.1994.1594. [DOI] [PubMed] [Google Scholar]
  52. Zorzato F., Fujii J., Otsu K., Phillips M., Green N. M., Lai F. A., Meissner G., MacLennan D. H. Molecular cloning of cDNA encoding human and rabbit forms of the Ca2+ release channel (ryanodine receptor) of skeletal muscle sarcoplasmic reticulum. J Biol Chem. 1990 Feb 5;265(4):2244–2256. [PubMed] [Google Scholar]
  53. van Heel M., Harauz G., Orlova E. V., Schmidt R., Schatz M. A new generation of the IMAGIC image processing system. J Struct Biol. 1996 Jan-Feb;116(1):17–24. doi: 10.1006/jsbi.1996.0004. [DOI] [PubMed] [Google Scholar]

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