Triadin binding to the C-terminal luminal loop of the ryanodine receptor is important for skeletal muscle excitation contraction coupling

Sanjeewa A Goonasekera, Nicole A Beard, Linda Groom, Takashi Kimura, Alla D Lyfenko, Andrew Rosenfeld, Isabelle Marty, Angela F. Dulhunty, Robert T Dirksen

    Research output: Contribution to journalArticle

    48 Citations (Scopus)

    Abstract

    Ca(2+) release from intracellular stores is controlled by complex interactions between multiple proteins. Triadin is a transmembrane glycoprotein of the junctional sarcoplasmic reticulum of striated muscle that interacts with both calsequestrin and the type 1 ryanodine receptor (RyR1) to communicate changes in luminal Ca(2+) to the release machinery. However, the potential impact of the triadin association with RyR1 in skeletal muscle excitation-contraction coupling remains elusive. Here we show that triadin binding to RyR1 is critically important for rapid Ca(2+) release during excitation-contraction coupling. To assess the functional impact of the triadin-RyR1 interaction, we expressed RyR1 mutants in which one or more of three negatively charged residues (D4878, D4907, and E4908) in the terminal RyR1 intraluminal loop were mutated to alanines in RyR1-null (dyspedic) myotubes. Coimmunoprecipitation revealed that triadin, but not junctin, binding to RyR1 was abolished in the triple (D4878A/D4907A/E4908A) mutant and one of the double (D4907A/E4908A) mutants, partially reduced in the D4878A/D4907A double mutant, but not affected by either individual (D4878A, D4907A, E4908A) mutations or the D4878A/E4908A double mutation. Functional studies revealed that the rate of voltage- and ligand-gated SR Ca(2+) release were reduced in proportion to the degree of interruption in triadin binding. Ryanodine binding, single channel recording, and calcium release experiments conducted on WT and triple mutant channels in the absence of triadin demonstrated that the luminal loop mutations do not directly alter RyR1 function. These findings demonstrate that junctin and triadin bind to different sites on RyR1 and that triadin plays an important role in ensuring rapid Ca(2+) release during excitation-contraction coupling in skeletal muscle.

    Original languageEnglish
    Pages (from-to)365-378
    Number of pages14
    JournalJournal of General Physiology
    Volume130
    Issue number4
    DOIs
    Publication statusPublished - Oct 2007

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    Excitation Contraction Coupling
    Ryanodine Receptor Calcium Release Channel
    Muscle Contraction
    Skeletal Muscle
    Mutation
    triadin
    Calsequestrin
    Ryanodine
    Striated Muscle
    Skeletal Muscle Fibers
    Sarcoplasmic Reticulum
    Calcium Channels
    Alanine
    Glycoproteins

    Cite this

    Goonasekera, Sanjeewa A ; Beard, Nicole A ; Groom, Linda ; Kimura, Takashi ; Lyfenko, Alla D ; Rosenfeld, Andrew ; Marty, Isabelle ; Dulhunty, Angela F. ; Dirksen, Robert T. / Triadin binding to the C-terminal luminal loop of the ryanodine receptor is important for skeletal muscle excitation contraction coupling. In: Journal of General Physiology. 2007 ; Vol. 130, No. 4. pp. 365-378.
    @article{f2a34bf7a02247cd8622318a102d21c5,
    title = "Triadin binding to the C-terminal luminal loop of the ryanodine receptor is important for skeletal muscle excitation contraction coupling",
    abstract = "Ca(2+) release from intracellular stores is controlled by complex interactions between multiple proteins. Triadin is a transmembrane glycoprotein of the junctional sarcoplasmic reticulum of striated muscle that interacts with both calsequestrin and the type 1 ryanodine receptor (RyR1) to communicate changes in luminal Ca(2+) to the release machinery. However, the potential impact of the triadin association with RyR1 in skeletal muscle excitation-contraction coupling remains elusive. Here we show that triadin binding to RyR1 is critically important for rapid Ca(2+) release during excitation-contraction coupling. To assess the functional impact of the triadin-RyR1 interaction, we expressed RyR1 mutants in which one or more of three negatively charged residues (D4878, D4907, and E4908) in the terminal RyR1 intraluminal loop were mutated to alanines in RyR1-null (dyspedic) myotubes. Coimmunoprecipitation revealed that triadin, but not junctin, binding to RyR1 was abolished in the triple (D4878A/D4907A/E4908A) mutant and one of the double (D4907A/E4908A) mutants, partially reduced in the D4878A/D4907A double mutant, but not affected by either individual (D4878A, D4907A, E4908A) mutations or the D4878A/E4908A double mutation. Functional studies revealed that the rate of voltage- and ligand-gated SR Ca(2+) release were reduced in proportion to the degree of interruption in triadin binding. Ryanodine binding, single channel recording, and calcium release experiments conducted on WT and triple mutant channels in the absence of triadin demonstrated that the luminal loop mutations do not directly alter RyR1 function. These findings demonstrate that junctin and triadin bind to different sites on RyR1 and that triadin plays an important role in ensuring rapid Ca(2+) release during excitation-contraction coupling in skeletal muscle.",
    keywords = "Amino Acid Substitution, Animals, Binding Sites, Calcium Channels, Calcium Channels, L-Type, Calcium Signaling, Calcium-Binding Proteins, Calsequestrin, Carrier Proteins, Cell Line, Transformed, Cells, Cultured, Electrophysiology, Kinetics, Membrane Proteins, Mice, Mixed Function Oxygenases, Models, Biological, Muscle Contraction, Muscle Fibers, Skeletal, Muscle Proteins, Protein Binding, Protein Interaction Mapping, Rabbits, Ryanodine Receptor Calcium Release Channel, Sarcoplasmic Reticulum",
    author = "Goonasekera, {Sanjeewa A} and Beard, {Nicole A} and Linda Groom and Takashi Kimura and Lyfenko, {Alla D} and Andrew Rosenfeld and Isabelle Marty and Dulhunty, {Angela F.} and Dirksen, {Robert T}",
    year = "2007",
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    doi = "10.1085/jgp.200709790",
    language = "English",
    volume = "130",
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    Goonasekera, SA, Beard, NA, Groom, L, Kimura, T, Lyfenko, AD, Rosenfeld, A, Marty, I, Dulhunty, AF & Dirksen, RT 2007, 'Triadin binding to the C-terminal luminal loop of the ryanodine receptor is important for skeletal muscle excitation contraction coupling', Journal of General Physiology, vol. 130, no. 4, pp. 365-378. https://doi.org/10.1085/jgp.200709790

    Triadin binding to the C-terminal luminal loop of the ryanodine receptor is important for skeletal muscle excitation contraction coupling. / Goonasekera, Sanjeewa A; Beard, Nicole A; Groom, Linda; Kimura, Takashi; Lyfenko, Alla D; Rosenfeld, Andrew; Marty, Isabelle; Dulhunty, Angela F.; Dirksen, Robert T.

    In: Journal of General Physiology, Vol. 130, No. 4, 10.2007, p. 365-378.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Triadin binding to the C-terminal luminal loop of the ryanodine receptor is important for skeletal muscle excitation contraction coupling

    AU - Goonasekera, Sanjeewa A

    AU - Beard, Nicole A

    AU - Groom, Linda

    AU - Kimura, Takashi

    AU - Lyfenko, Alla D

    AU - Rosenfeld, Andrew

    AU - Marty, Isabelle

    AU - Dulhunty, Angela F.

    AU - Dirksen, Robert T

    PY - 2007/10

    Y1 - 2007/10

    N2 - Ca(2+) release from intracellular stores is controlled by complex interactions between multiple proteins. Triadin is a transmembrane glycoprotein of the junctional sarcoplasmic reticulum of striated muscle that interacts with both calsequestrin and the type 1 ryanodine receptor (RyR1) to communicate changes in luminal Ca(2+) to the release machinery. However, the potential impact of the triadin association with RyR1 in skeletal muscle excitation-contraction coupling remains elusive. Here we show that triadin binding to RyR1 is critically important for rapid Ca(2+) release during excitation-contraction coupling. To assess the functional impact of the triadin-RyR1 interaction, we expressed RyR1 mutants in which one or more of three negatively charged residues (D4878, D4907, and E4908) in the terminal RyR1 intraluminal loop were mutated to alanines in RyR1-null (dyspedic) myotubes. Coimmunoprecipitation revealed that triadin, but not junctin, binding to RyR1 was abolished in the triple (D4878A/D4907A/E4908A) mutant and one of the double (D4907A/E4908A) mutants, partially reduced in the D4878A/D4907A double mutant, but not affected by either individual (D4878A, D4907A, E4908A) mutations or the D4878A/E4908A double mutation. Functional studies revealed that the rate of voltage- and ligand-gated SR Ca(2+) release were reduced in proportion to the degree of interruption in triadin binding. Ryanodine binding, single channel recording, and calcium release experiments conducted on WT and triple mutant channels in the absence of triadin demonstrated that the luminal loop mutations do not directly alter RyR1 function. These findings demonstrate that junctin and triadin bind to different sites on RyR1 and that triadin plays an important role in ensuring rapid Ca(2+) release during excitation-contraction coupling in skeletal muscle.

    AB - Ca(2+) release from intracellular stores is controlled by complex interactions between multiple proteins. Triadin is a transmembrane glycoprotein of the junctional sarcoplasmic reticulum of striated muscle that interacts with both calsequestrin and the type 1 ryanodine receptor (RyR1) to communicate changes in luminal Ca(2+) to the release machinery. However, the potential impact of the triadin association with RyR1 in skeletal muscle excitation-contraction coupling remains elusive. Here we show that triadin binding to RyR1 is critically important for rapid Ca(2+) release during excitation-contraction coupling. To assess the functional impact of the triadin-RyR1 interaction, we expressed RyR1 mutants in which one or more of three negatively charged residues (D4878, D4907, and E4908) in the terminal RyR1 intraluminal loop were mutated to alanines in RyR1-null (dyspedic) myotubes. Coimmunoprecipitation revealed that triadin, but not junctin, binding to RyR1 was abolished in the triple (D4878A/D4907A/E4908A) mutant and one of the double (D4907A/E4908A) mutants, partially reduced in the D4878A/D4907A double mutant, but not affected by either individual (D4878A, D4907A, E4908A) mutations or the D4878A/E4908A double mutation. Functional studies revealed that the rate of voltage- and ligand-gated SR Ca(2+) release were reduced in proportion to the degree of interruption in triadin binding. Ryanodine binding, single channel recording, and calcium release experiments conducted on WT and triple mutant channels in the absence of triadin demonstrated that the luminal loop mutations do not directly alter RyR1 function. These findings demonstrate that junctin and triadin bind to different sites on RyR1 and that triadin plays an important role in ensuring rapid Ca(2+) release during excitation-contraction coupling in skeletal muscle.

    KW - Amino Acid Substitution

    KW - Animals

    KW - Binding Sites

    KW - Calcium Channels

    KW - Calcium Channels, L-Type

    KW - Calcium Signaling

    KW - Calcium-Binding Proteins

    KW - Calsequestrin

    KW - Carrier Proteins

    KW - Cell Line, Transformed

    KW - Cells, Cultured

    KW - Electrophysiology

    KW - Kinetics

    KW - Membrane Proteins

    KW - Mice

    KW - Mixed Function Oxygenases

    KW - Models, Biological

    KW - Muscle Contraction

    KW - Muscle Fibers, Skeletal

    KW - Muscle Proteins

    KW - Protein Binding

    KW - Protein Interaction Mapping

    KW - Rabbits

    KW - Ryanodine Receptor Calcium Release Channel

    KW - Sarcoplasmic Reticulum

    UR - http://jgp.rupress.org/content/130/4/365

    U2 - 10.1085/jgp.200709790

    DO - 10.1085/jgp.200709790

    M3 - Article

    VL - 130

    SP - 365

    EP - 378

    JO - Journal of General Physiology

    JF - Journal of General Physiology

    SN - 0022-1295

    IS - 4

    ER -