Interdependence of ryanodine binding, oligomeric receptor interactions, and Ca2+ release regulation in junctional sarcoplasmic reticulum

https://doi.org/10.1016/0003-9861(91)90615-PGet rights and content

Abstract

We have examined ryanodine binding to its receptor (RR) and compared its effect on Ca2+ release to the Ca2+ release triggered by Ca2+ plus ATP, using vesicular fragments of junctional terminal cisternae (JTC) obtained from skeletal muscle. Ryanodine binding is slow (taking hours or days to complete) and is highly temperature (Q10 = 4) and Ca2+ dependent. At equilibrium, the extent of binding increases as the concentration of ryanodine is raised above 10−9m, exhibiting negative cooperativity and reaching the stoichiometry of the 560,000-Da RR chains near 10−5m ryanodine. The specificity of the high affinity binding is demonstrated by competitive binding of ryanodine analogs. Kinetic studies using rapid filtration show that, in the absence of ryanodine, rapid (k = 15 s−1) release of Ca2+ follows a triggering exposure of loaded JTC vesicles to perfusion media containing Ca2+ plus ATP. Induction of this release has no lag period and displays minimal temperature dependence. In contrast, prolonged exposure of JTC vesicles to low (10−7m) ryanodine concentrations changes the JTC to a state permitting slow (k = 1 s−1) release of Ca2+ even in the absence of the Ca2+ plus ATP trigger. Higher (>μm) concentrations of ryanodine do not allow any Ca2+ release and prevent even the release normally triggered by Ca2+ plus ATP. Our data suggest that ryanodine binds to the open state of the tetrameric RR, inducing protein conformational changes and altered oligomeric interactions. Binding of the first molecule of ryanodine to one of the four binding sites on the receptor produces a partially closed and low conductance state of the Ca2+ release channel and reduces the ryanodine binding affinity of the remaining sites. Ryanodine occupancy of all four binding sites on the receptor completes closure of the Ca2+ channel and blocks the triggering action of Ca2+ plus ATP. The tetrameric association of the RR chains is demonstrated by crosslinking with bifunctional reagents, generating crosslinked tetramers that retain ryanodine binding and Ca2+ release functions.

References (67)

  • J.S. Smith et al.

    Biophys. J

    (1986)
  • T. Imagawa et al.

    J. Biol. Chem

    (1987)
  • G. Meissner

    Biochim. Biophys. Acta

    (1975)
  • K.P. Campbell et al.

    J. Biol. Chem

    (1987)
  • F. Zorzato et al.

    J. Biol. Chem

    (1990)
  • E. Rousseau et al.

    Biophys. J

    (1986)
  • N. Ikemoto et al.

    J. Biol. Chem

    (1985)
  • G. Salama et al.

    J. Biol. Chem

    (1984)
  • P. Palade

    J. Biol. Chem

    (1987)
  • J.L. Trimm et al.

    J. Biol. Chem

    (1986)
  • J.J. Abramson et al.

    J. Biol. Chem

    (1988)
  • J.J. Abramson et al.

    Arch. Biochem. Biophys

    (1988)
  • N.F. Zaidi et al.

    J. Biol. Chem

    (1989)
  • B.K. Chamberlain et al.

    J. Biol. Chem

    (1983)
  • S. Seiler et al.

    J. Biol. Chem

    (1984)
  • B.H. Alderson et al.

    Biochim. Biophys. Acta

    (1987)
  • M. Inui et al.

    J. Biol. Chem

    (1988)
  • M. Michalak et al.

    Biochim. Biophys. Acta

    (1988)
  • O.H. Lowry et al.

    J. Biol. Chem

    (1951)
  • M. Moutin et al.

    J. Biol. Chem

    (1988)
  • G. Meissner et al.

    J. Biol. Chem

    (1989)
  • G. Meissner et al.

    J. Biol. Chem

    (1987)
  • F.A. Lattanzio et al.

    J. Biol. Chem

    (1987)
  • J.A. Airey et al.

    J. Biol. Chem

    (1990)
  • K. Nagasaki et al.

    Cell Calcium

    (1989)
  • J.J. Feher et al.

    Biochim. Biophys. Acta

    (1985)
  • C. Franzini-Armstrong

    J. Cell Biol

    (1970)
  • A. Saito et al.

    J. Cell Biol

    (1984)
  • R.M. Kawamoto et al.

    J. Cell Biol

    (1986)
  • J.S. Smith et al.

    Nature

    (1985)
  • A. Fabiato et al.

    J. Physiol. (London)

    (1975)
  • N. Yamamoto et al.

    J. Biochem. (Tokyo)

    (1982)
  • G. Meissner et al.

    Biochemistry

    (1986)
  • Cited by (47)

    • Structural Basis for Gating and Activation of RyR1

      2016, Cell
      Citation Excerpt :

      Increased ryanoid concentrations (∼100 μM) lock the channel in a non-conducting state (Zimányi et al., 1992). RyRs possess one high-affinity ryanodine-binding site per tetramer, and up to three lower-affinity sites (Carroll et al., 1991). Several lines of evidence suggest that the high-affinity ryanodine-binding site is located within the transmembrane pore of the channel.

    • Inhibitory ryanodine prevents ryanodine receptor-mediated Ca<sup>2+</sup> release without affecting endoplasmic reticulum Ca<sup>2+</sup> content in primary hippocampal neurons

      2015, Biochemical and Biophysical Research Communications
      Citation Excerpt :

      Mounting evidence also points to a significant role of RyR-mediated Ca2+ release on normal [7,8] and pathological neuronal function [5,9]. Ryanodine is a cell permeant plant alkaloid [10] that binds selectively and with high affinity to the RyR channel protein but dissociates very slowly once bound [11]. These binding properties allowed the initial identification of the RyR protein (reviewed in Ref. [12]).

    • RyR1 S-Nitrosylation Underlies Environmental Heat Stroke and Sudden Death in Y522S RyR1 Knockin Mice

      2008, Cell
      Citation Excerpt :

      The previously described binding assays were performed at room temperature. RyR1 is, however, not stable for extended periods of time at higher temperatures (Carroll et al., 1991), making equilibrium binding studies at physiologic temperatures difficult. To circumvent this problem, we assessed the rate of association of [3H] ryanodine to skeletal muscle membranes from RyR1Y522S/wt and RyR1wt/wt mice at different temperatures and in the presence or absence of either DTT or AA.

    • Effects of quercetin on single Ca<sup>2+</sup> release channel behavior of skeletal muscle

      2002, Biophysical Journal
      Citation Excerpt :

      An analysis of Po and the gating mode of single CRC showed that the mean open-time and closed-time constants (τo and τc) and their proportions were significantly different for the caffeine- and quercetin-activated CRCs, even though Po values were similar (Table 2). Ryanodine binding to its high-affinity site(s) stabilizes the open state of CRC, however, open channel conductance is subnormal (Rousseau et al., 1987; Carroll et al., 1991; Pessah and Zimanyi, 1991; Buck et al., 1992). The I-V relations in Fig. 3 D show that the conductances and levels of subconductance state formed by 10 μM ryanodine were similar in all cases (Fig. 3), suggesting that quercetin does not modify the ion selectivity of the CRC.

    View all citing articles on Scopus

    This work was supported by grants from the National Institutes of Health (HL-27867) and the Muscular Dystrophy Association.

    View full text