Regulation of Ryanodine receptors by Calsequestrin: Effect of high luminal Ca2+ and phosphorylation

Nicole A Beard, Marco G Casarotto, Lan Wei, Magdolna Varsányi, Derek R Laver, Angela F. Dulhunty

    Research output: Contribution to journalArticle

    80 Citations (Scopus)

    Abstract

    Calsequestrin, the major calcium sequestering protein in the sarcoplasmic reticulum of muscle, forms a quaternary complex with the ryanodine receptor calcium release channel and the intrinsic membrane proteins triadin and junctin. We have investigated the possibility that calsequestrin is a luminal calcium concentration sensor for the ryanodine receptor. We measured the luminal calcium concentration at which calsequestrin dissociates from the ryanodine receptor and the effect of calsequestrin on the response of the ryanodine receptor to changes in luminal calcium. We provide electrophysiological and biochemical evidence that: 1), luminal calcium concentration of >/=4 mM dissociates calsequestrin from junctional face membrane, whereas in the range of 1-3 mM calsequestrin remains attached; 2), the association with calsequestrin inhibits ryanodine receptor activity, but amplifies its response to changes in luminal calcium concentration; and 3), under physiological calcium conditions (1 mM), phosphorylation of calsequestrin does not alter its ability to inhibit native ryanodine receptor activity when the anchoring proteins triadin and junctin are present. These data suggest that the quaternary complex is intact in vivo, and provides further evidence that calsequestrin is involved in the sarcoplasmic reticulum calcium signaling pathway and has a role as a luminal calcium sensor for the ryanodine receptor.

    Original languageEnglish
    Pages (from-to)3444-3454
    Number of pages11
    JournalBiophysical Journal
    Volume88
    Issue number5
    DOIs
    Publication statusPublished - May 2005

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    Calsequestrin
    Ryanodine Receptor Calcium Release Channel
    Phosphorylation
    Calcium
    Sarcoplasmic Reticulum
    Calcium Signaling
    Membrane Proteins
    Proteins

    Cite this

    Beard, Nicole A ; Casarotto, Marco G ; Wei, Lan ; Varsányi, Magdolna ; Laver, Derek R ; Dulhunty, Angela F. / Regulation of Ryanodine receptors by Calsequestrin: Effect of high luminal Ca2+ and phosphorylation. In: Biophysical Journal. 2005 ; Vol. 88, No. 5. pp. 3444-3454.
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    abstract = "Calsequestrin, the major calcium sequestering protein in the sarcoplasmic reticulum of muscle, forms a quaternary complex with the ryanodine receptor calcium release channel and the intrinsic membrane proteins triadin and junctin. We have investigated the possibility that calsequestrin is a luminal calcium concentration sensor for the ryanodine receptor. We measured the luminal calcium concentration at which calsequestrin dissociates from the ryanodine receptor and the effect of calsequestrin on the response of the ryanodine receptor to changes in luminal calcium. We provide electrophysiological and biochemical evidence that: 1), luminal calcium concentration of >/=4 mM dissociates calsequestrin from junctional face membrane, whereas in the range of 1-3 mM calsequestrin remains attached; 2), the association with calsequestrin inhibits ryanodine receptor activity, but amplifies its response to changes in luminal calcium concentration; and 3), under physiological calcium conditions (1 mM), phosphorylation of calsequestrin does not alter its ability to inhibit native ryanodine receptor activity when the anchoring proteins triadin and junctin are present. These data suggest that the quaternary complex is intact in vivo, and provides further evidence that calsequestrin is involved in the sarcoplasmic reticulum calcium signaling pathway and has a role as a luminal calcium sensor for the ryanodine receptor.",
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    Regulation of Ryanodine receptors by Calsequestrin: Effect of high luminal Ca2+ and phosphorylation. / Beard, Nicole A; Casarotto, Marco G; Wei, Lan; Varsányi, Magdolna; Laver, Derek R; Dulhunty, Angela F.

    In: Biophysical Journal, Vol. 88, No. 5, 05.2005, p. 3444-3454.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Regulation of Ryanodine receptors by Calsequestrin: Effect of high luminal Ca2+ and phosphorylation

    AU - Beard, Nicole A

    AU - Casarotto, Marco G

    AU - Wei, Lan

    AU - Varsányi, Magdolna

    AU - Laver, Derek R

    AU - Dulhunty, Angela F.

    PY - 2005/5

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    N2 - Calsequestrin, the major calcium sequestering protein in the sarcoplasmic reticulum of muscle, forms a quaternary complex with the ryanodine receptor calcium release channel and the intrinsic membrane proteins triadin and junctin. We have investigated the possibility that calsequestrin is a luminal calcium concentration sensor for the ryanodine receptor. We measured the luminal calcium concentration at which calsequestrin dissociates from the ryanodine receptor and the effect of calsequestrin on the response of the ryanodine receptor to changes in luminal calcium. We provide electrophysiological and biochemical evidence that: 1), luminal calcium concentration of >/=4 mM dissociates calsequestrin from junctional face membrane, whereas in the range of 1-3 mM calsequestrin remains attached; 2), the association with calsequestrin inhibits ryanodine receptor activity, but amplifies its response to changes in luminal calcium concentration; and 3), under physiological calcium conditions (1 mM), phosphorylation of calsequestrin does not alter its ability to inhibit native ryanodine receptor activity when the anchoring proteins triadin and junctin are present. These data suggest that the quaternary complex is intact in vivo, and provides further evidence that calsequestrin is involved in the sarcoplasmic reticulum calcium signaling pathway and has a role as a luminal calcium sensor for the ryanodine receptor.

    AB - Calsequestrin, the major calcium sequestering protein in the sarcoplasmic reticulum of muscle, forms a quaternary complex with the ryanodine receptor calcium release channel and the intrinsic membrane proteins triadin and junctin. We have investigated the possibility that calsequestrin is a luminal calcium concentration sensor for the ryanodine receptor. We measured the luminal calcium concentration at which calsequestrin dissociates from the ryanodine receptor and the effect of calsequestrin on the response of the ryanodine receptor to changes in luminal calcium. We provide electrophysiological and biochemical evidence that: 1), luminal calcium concentration of >/=4 mM dissociates calsequestrin from junctional face membrane, whereas in the range of 1-3 mM calsequestrin remains attached; 2), the association with calsequestrin inhibits ryanodine receptor activity, but amplifies its response to changes in luminal calcium concentration; and 3), under physiological calcium conditions (1 mM), phosphorylation of calsequestrin does not alter its ability to inhibit native ryanodine receptor activity when the anchoring proteins triadin and junctin are present. These data suggest that the quaternary complex is intact in vivo, and provides further evidence that calsequestrin is involved in the sarcoplasmic reticulum calcium signaling pathway and has a role as a luminal calcium sensor for the ryanodine receptor.

    KW - Acid Phosphatase

    KW - Animals

    KW - Calcium

    KW - Calcium-Binding Proteins

    KW - Calsequestrin

    KW - Carrier Proteins

    KW - Casein Kinase II

    KW - Chromatography

    KW - Dose-Response Relationship, Drug

    KW - Electrophoresis, Polyacrylamide Gel

    KW - Electrophysiology

    KW - Glutathione

    KW - Immunoblotting

    KW - Lipid Bilayers

    KW - Magnetic Resonance Spectroscopy

    KW - Membrane Proteins

    KW - Mixed Function Oxygenases

    KW - Muscle Proteins

    KW - Muscle, Skeletal

    KW - Muscles

    KW - Phosphorylation

    KW - Protein Conformation

    KW - Rabbits

    KW - Recombinant Fusion Proteins

    KW - Recombinant Proteins

    KW - Ryanodine Receptor Calcium Release Channel

    KW - Sarcoplasmic Reticulum

    KW - Signal Transduction

    U2 - 10.1529/biophysj.104.051441

    DO - 10.1529/biophysj.104.051441

    M3 - Article

    VL - 88

    SP - 3444

    EP - 3454

    JO - Biophysical Journal

    JF - Biophysical Journal

    SN - 0006-3495

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