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. 1999 Jul;77(1):189–203. doi: 10.1016/S0006-3495(99)76881-5

Activation and inhibition of skeletal RyR channels by a part of the skeletal DHPR II-III loop: effects of DHPR Ser687 and FKBP12.

A F Dulhunty 1, D R Laver 1, E M Gallant 1, M G Casarotto 1, S M Pace 1, S Curtis 1
PMCID: PMC1300321  PMID: 10388749

Abstract

Peptides, corresponding to sequences in the N-terminal region of the skeletal muscle dihydropyridine receptor (DHPR) II-III loop, have been tested on sarcoplasmic reticulum (SR) Ca2+ release and ryanodine receptor (RyR) activity. The peptides were: A1, Thr671-Leu690; A2, Thr671-Leu690 with Ser687 Ala substitution; NB, Gly689-Lys708 and A1S, scrambled A1 sequence. The relative rates of peptide-induced Ca2+ release from normal (FKBP12+) SR were A2 > A1 > A1S > NB. Removal of FKBP12 reduced the rate of A1-induced Ca2+ release by approximately 30%. A1 and A2 (but not NB or A1S), in the cytoplasmic (cis) solution, either activated or inhibited single FKBP12+ RyRs. Maximum activation was seen at -40 mV, with 10 microM A1 or 50 nM A2. The greatest A1-induced increase in mean current (sixfold) was seen with 100 nM cis Ca2+. Inhibition by A1 was greatest at +40 mV (or when permeant ions flowed from cytoplasm to SR lumen) with 100 microM cis Ca2+, where channel activity was almost fully inhibited. A1 did not activate FKBP12-stripped RyRs, although peptide-induced inhibition remained. The results show that peptide A activation of RyRs does not require DHPR Ser687, but required FKBP12 binding to RyRs. Peptide A must interact with different sites to activate or inhibit RyRs, because current direction-, voltage-, cis [Ca2+]-, and FKBP12-dependence of activation and inhibition differ.

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Selected References

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  1. Ahern G. P., Junankar P. R., Dulhunty A. F. Ryanodine receptors from rabbit skeletal muscle are reversibly activated by rapamycin. Neurosci Lett. 1997 Apr 4;225(2):81–84. doi: 10.1016/s0304-3940(97)00193-6. [DOI] [PubMed] [Google Scholar]
  2. Ahern G. P., Junankar P. R., Dulhunty A. F. Single channel activity of the ryanodine receptor calcium release channel is modulated by FK-506. FEBS Lett. 1994 Oct 3;352(3):369–374. doi: 10.1016/0014-5793(94)01001-3. [DOI] [PubMed] [Google Scholar]
  3. Ahern G. P., Junankar P. R., Dulhunty A. F. Subconductance states in single-channel activity of skeletal muscle ryanodine receptors after removal of FKBP12. Biophys J. 1997 Jan;72(1):146–162. doi: 10.1016/S0006-3495(97)78654-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Brillantes A. B., Ondrias K., Scott A., Kobrinsky E., Ondriasová E., Moschella M. C., Jayaraman T., Landers M., Ehrlich B. E., Marks A. R. Stabilization of calcium release channel (ryanodine receptor) function by FK506-binding protein. Cell. 1994 May 20;77(4):513–523. doi: 10.1016/0092-8674(94)90214-3. [DOI] [PubMed] [Google Scholar]
  6. Cameron A. M., Nucifora F. C., Jr, Fung E. T., Livingston D. J., Aldape R. A., Ross C. A., Snyder S. H. FKBP12 binds the inositol 1,4,5-trisphosphate receptor at leucine-proline (1400-1401) and anchors calcineurin to this FK506-like domain. J Biol Chem. 1997 Oct 31;272(44):27582–27588. doi: 10.1074/jbc.272.44.27582. [DOI] [PubMed] [Google Scholar]
  7. El-Hayek R., Ikemoto N. Identification of the minimum essential region in the II-III loop of the dihydropyridine receptor alpha 1 subunit required for activation of skeletal muscle-type excitation-contraction coupling. Biochemistry. 1998 May 12;37(19):7015–7020. doi: 10.1021/bi972907o. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Lamb G. D., Stephenson D. G. Effect of Mg2+ on the control of Ca2+ release in skeletal muscle fibres of the toad. J Physiol. 1991 Mar;434:507–528. doi: 10.1113/jphysiol.1991.sp018483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lamb G. D., Stephenson D. G. Effects of FK506 and rapamycin on excitation-contraction coupling in skeletal muscle fibres of the rat. J Physiol. 1996 Jul 15;494(Pt 2):569–576. doi: 10.1113/jphysiol.1996.sp021514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Laver D. R., Baynes T. M., Dulhunty A. F. Magnesium inhibition of ryanodine-receptor calcium channels: evidence for two independent mechanisms. J Membr Biol. 1997 Apr 1;156(3):213–229. doi: 10.1007/s002329900202. [DOI] [PubMed] [Google Scholar]
  12. Laver D. R., Owen V. J., Junankar P. R., Taske N. L., Dulhunty A. F., Lamb G. D. Reduced inhibitory effect of Mg2+ on ryanodine receptor-Ca2+ release channels in malignant hyperthermia. Biophys J. 1997 Oct;73(4):1913–1924. doi: 10.1016/S0006-3495(97)78222-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Laver D. R., Roden L. D., Ahern G. P., Eager K. R., Junankar P. R., Dulhunty A. F. Cytoplasmic Ca2+ inhibits the ryanodine receptor from cardiac muscle. J Membr Biol. 1995 Sep;147(1):7–22. doi: 10.1007/BF00235394. [DOI] [PubMed] [Google Scholar]
  14. Leong P., MacLennan D. H. A 37-amino acid sequence in the skeletal muscle ryanodine receptor interacts with the cytoplasmic loop between domains II and III in the skeletal muscle dihydropyridine receptor. J Biol Chem. 1998 Apr 3;273(14):7791–7794. doi: 10.1074/jbc.273.14.7791. [DOI] [PubMed] [Google Scholar]
  15. Lu X., Xu L., Meissner G. Activation of the skeletal muscle calcium release channel by a cytoplasmic loop of the dihydropyridine receptor. J Biol Chem. 1994 Mar 4;269(9):6511–6516. [PubMed] [Google Scholar]
  16. Lu X., Xu L., Meissner G. Phosphorylation of dihydropyridine receptor II-III loop peptide regulates skeletal muscle calcium release channel function. Evidence for an essential role of the beta-OH group of Ser687. J Biol Chem. 1995 Aug 4;270(31):18459–18464. doi: 10.1074/jbc.270.31.18459. [DOI] [PubMed] [Google Scholar]
  17. MacKinnon R., Miller C. Mechanism of charybdotoxin block of the high-conductance, Ca2+-activated K+ channel. J Gen Physiol. 1988 Mar;91(3):335–349. doi: 10.1085/jgp.91.3.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marty I., Robert M., Villaz M., De Jongh K., Lai Y., Catterall W. A., Ronjat M. Biochemical evidence for a complex involving dihydropyridine receptor and ryanodine receptor in triad junctions of skeletal muscle. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2270–2274. doi: 10.1073/pnas.91.6.2270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mayrleitner M., Timerman A. P., Wiederrecht G., Fleischer S. The calcium release channel of sarcoplasmic reticulum is modulated by FK-506 binding protein: effect of FKBP-12 on single channel activity of the skeletal muscle ryanodine receptor. Cell Calcium. 1994 Feb;15(2):99–108. doi: 10.1016/0143-4160(94)90048-5. [DOI] [PubMed] [Google Scholar]
  20. Mead F. C., Sullivan D., Williams A. J. Evidence for negative charge in the conduction pathway of the cardiac ryanodine receptor channel provided by the interaction of K+ channel N-type inactivation peptides. J Membr Biol. 1998 Jun 1;163(3):225–234. doi: 10.1007/s002329900386. [DOI] [PubMed] [Google Scholar]
  21. Melzer W., Rios E., Schneider M. F. A general procedure for determining the rate of calcium release from the sarcoplasmic reticulum in skeletal muscle fibers. Biophys J. 1987 Jun;51(6):849–863. doi: 10.1016/S0006-3495(87)83413-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Murray B. E., Ohlendieck K. Cross-linking analysis of the ryanodine receptor and alpha1-dihydropyridine receptor in rabbit skeletal muscle triads. Biochem J. 1997 Jun 1;324(Pt 2):689–696. doi: 10.1042/bj3240689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nakai J., Sekiguchi N., Rando T. A., Allen P. D., Beam K. G. Two regions of the ryanodine receptor involved in coupling with L-type Ca2+ channels. J Biol Chem. 1998 May 29;273(22):13403–13406. doi: 10.1074/jbc.273.22.13403. [DOI] [PubMed] [Google Scholar]
  24. Nakai J., Tanabe T., Konno T., Adams B., Beam K. G. Localization in the II-III loop of the dihydropyridine receptor of a sequence critical for excitation-contraction coupling. J Biol Chem. 1998 Sep 25;273(39):24983–24986. doi: 10.1074/jbc.273.39.24983. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Sagara Y., Inesi G. Inhibition of the sarcoplasmic reticulum Ca2+ transport ATPase by thapsigargin at subnanomolar concentrations. J Biol Chem. 1991 Jul 25;266(21):13503–13506. [PubMed] [Google Scholar]
  28. 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]
  29. Tanabe T., Beam K. G., Adams B. A., Niidome T., Numa S. Regions of the skeletal muscle dihydropyridine receptor critical for excitation-contraction coupling. Nature. 1990 Aug 9;346(6284):567–569. doi: 10.1038/346567a0. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. 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]
  32. Wagenknecht T., Grassucci R., Berkowitz J., Wiederrecht G. J., Xin H. B., Fleischer S. Cryoelectron microscopy resolves FK506-binding protein sites on the skeletal muscle ryanodine receptor. Biophys J. 1996 Apr;70(4):1709–1715. doi: 10.1016/S0006-3495(96)79733-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wagenknecht T., Radermacher M., Grassucci R., Berkowitz J., Xin H. B., Fleischer S. Locations of calmodulin and FK506-binding protein on the three-dimensional architecture of the skeletal muscle ryanodine receptor. J Biol Chem. 1997 Dec 19;272(51):32463–32471. doi: 10.1074/jbc.272.51.32463. [DOI] [PubMed] [Google Scholar]
  34. Wagenknecht T., Radermacher M. Ryanodine receptors: structure and macromolecular interactions. Curr Opin Struct Biol. 1997 Apr;7(2):258–265. doi: 10.1016/s0959-440x(97)80034-6. [DOI] [PubMed] [Google Scholar]
  35. Xu L., Mann G., Meissner G. Regulation of cardiac Ca2+ release channel (ryanodine receptor) by Ca2+, H+, Mg2+, and adenine nucleotides under normal and simulated ischemic conditions. Circ Res. 1996 Dec;79(6):1100–1109. doi: 10.1161/01.res.79.6.1100. [DOI] [PubMed] [Google Scholar]
  36. Yamazawa T., Takeshima H., Shimuta M., Iino M. A region of the ryanodine receptor critical for excitation-contraction coupling in skeletal muscle. J Biol Chem. 1997 Mar 28;272(13):8161–8164. doi: 10.1074/jbc.272.13.8161. [DOI] [PubMed] [Google Scholar]
  37. el-Hayek R., Antoniu B., Wang J., Hamilton S. L., Ikemoto N. Identification of calcium release-triggering and blocking regions of the II-III loop of the skeletal muscle dihydropyridine receptor. J Biol Chem. 1995 Sep 22;270(38):22116–22118. doi: 10.1074/jbc.270.38.22116. [DOI] [PubMed] [Google Scholar]

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