Association of Triadin with the Ryanodine Receptor and Calsequestrin in the Lumen of the Sarcoplasmic Reticulum (*)

Triadin is a major membrane protein that is specifically localized in the junctional sarcoplasmic reticulum of skeletal muscle and is thought to play an important role in muscle excitation-contraction coupling. In order to identify the proteins in the skeletal muscle that interact with triadin, the cytoplasmic and luminal domains of triadin were expressed as glutathione S-transferase fusion proteins and immobilized to glutathione-Sepharose to form affinity columns. Using these affinity columns, we find that triadin binds specifically to the ryanodine receptor/Carelease channel and the Ca-binding protein calsequestrin from CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid)-solubilized skeletal muscle homogenates. The luminal but not the cytoplasmic domain of triadin-glutathione S-transferase fusion protein binds [3H]ryanodine receptor, whereas neither the cytoplasmic nor the luminal portion of triadin binds [3H]PN-200-100-labeled dihydropyridine receptor. In addition, the luminal domain of triadin interacts with calsequestrin in a Ca-dependent manner and is capable of inhibiting the reassociation of calsequestrin to the junctional face membrane. These results suggest that triadin is the previously unidentified transmembrane protein that anchors calsequestrin to the junctional region of the sarcoplasmic reticulum, and is involved in the functional coupling between calsequestrin and the ryanodine receptor/Carelease channel.

[1]  N. Thorn,et al.  Calcium/sodium exchange in purified secretory vesicles from bovine neurohypophyses. , 1983, Cell calcium.

[2]  S. Fleischer,et al.  Characterization of the junctional face membrane from terminal cisternae of sarcoplasmic reticulum , 1986, The Journal of cell biology.

[3]  M. Phillips,et al.  Molecular cloning of cDNA encoding human and rabbit forms of the Ca2+ release channel (ryanodine receptor) of skeletal muscle sarcoplasmic reticulum. , 1990, The Journal of biological chemistry.

[4]  Harold P. Erickson,et al.  Purification and reconstitution of the calcium release channel from skeletal muscle , 1988, Nature.

[5]  E. Ríos,et al.  Involvement of dihydropyridine receptors in excitation–contraction coupling in skeletal muscle , 1987, Nature.

[6]  K. Campbell,et al.  Solubilization and biochemical characterization of the high affinity [3H]ryanodine receptor from rabbit brain membranes. , 1990, The Journal of biological chemistry.

[7]  A. Belcastro,et al.  Intraluminal Ca2+ dependence of Ca2+ and ryanodine-mediated regulation of skeletal muscle sarcoplasmic reticulum Ca2+ release. , 1992, The Journal of biological chemistry.

[8]  C. Franzini-armstrong,et al.  The structure of calsequestrin in triads of vertebrate skeletal muscle: a deep-etch study , 1987, The Journal of cell biology.

[9]  M. Ronjat,et al.  Postulated role of calsequestrin in the regulation of calcium release from sarcoplasmic reticulum. , 1989, Biochemistry.

[10]  A. Caswell,et al.  Isolation, characterization, and localization of the spanning protein from skeletal muscle triads , 1986, The Journal of cell biology.

[11]  C. Slaughter,et al.  Primary structure and topological analysis of a skeletal muscle-specific junctional sarcoplasmic reticulum glycoprotein (triadin). , 1993, The Journal of biological chemistry.

[12]  L. Jones,et al.  Ca2+ binding effects on protein conformation and protein interactions of canine cardiac calsequestrin. , 1988, The Journal of biological chemistry.

[13]  K. Campbell,et al.  Identification and characterization of the high affinity [3H]ryanodine receptor of the junctional sarcoplasmic reticulum Ca2+ release channel. , 1987, The Journal of biological chemistry.

[14]  E. Delpont,et al.  [3H]nitrendipine receptors in skeletal muscle. , 1983, The Journal of biological chemistry.

[15]  K. Campbell,et al.  Subcellular distribution of the 1,4-dihydropyridine receptor in rabbit skeletal muscle in situ: an immunofluorescence and immunocolloidal gold- labeling study , 1989, The Journal of cell biology.

[16]  I. Pessah,et al.  Molecular interaction between ryanodine receptor and glycoprotein triadin involves redox cycling of functionally important hyperreactive sulfhydryls. , 1994, The Journal of biological chemistry.

[17]  K. Campbell,et al.  Biochemical characterization of ultrastructural localization of a major junctional sarcoplasmic reticulum glycoprotein (triadin). , 1993, The Journal of biological chemistry.

[18]  M. Kasai,et al.  Regulation of calcium channel in sarcoplasmic reticulum by calsequestrin. , 1994, Biochemical and biophysical research communications.

[19]  A. Caswell,et al.  Localization and partial characterization of the oligomeric disulfide-linked molecular weight 95,000 protein (triadin) which binds the ryanodine and dihydropyridine receptors in skeletal muscle triadic vesicles. , 1991, Biochemistry.

[20]  L. Jones,et al.  Rapid purification of calsequestrin from cardiac and skeletal muscle sarcoplasmic reticulum vesicles by Ca2+-dependent elution from phenyl-sepharose. , 1983, The Journal of biological chemistry.

[21]  M. Ronjat,et al.  Intravesicular calcium transient during calcium release from sarcoplasmic reticulum. , 1991, Biochemistry.

[22]  K. Campbell,et al.  Structural characterization of the 1,4-dihydropyridine receptor of the voltage-dependent Ca2+ channel from rabbit skeletal muscle. Evidence for two distinct high molecular weight subunits. , 1987, The Journal of biological chemistry.

[23]  K. Campbell,et al.  Specific absence of the alpha 1 subunit of the dihydropyridine receptor in mice with muscular dysgenesis. , 1989, The Journal of biological chemistry.

[24]  K. Campbell,et al.  Purified ryanodine receptor from skeletal muscle sarcoplasmic reticulum is the Ca2+-permeable pore of the calcium release channel. , 1987, The Journal of biological chemistry.

[25]  K. Campbell,et al.  The ryanodine receptor/Ca2+ release channel. , 1993, The Journal of biological chemistry.

[26]  B. Flucher,et al.  Localization of the α 1 and α 2 subunits of the dihydropyridine receptor and ankyrin in skeletal muscle triads , 1990, Neuron.

[27]  H. Takeshima,et al.  Primary structure and expression from complementary DNA of skeletal muscle ryanodine receptor , 1989, Nature.

[28]  P. Tempst,et al.  Surface topography analysis of the ryanodine receptor/junctional channel complex based on proteolysis sensitivity mapping. , 1990, The Journal of biological chemistry.

[29]  A. Dunker,et al.  Ca(2+)-induced folding and aggregation of skeletal muscle sarcoplasmic reticulum calsequestrin. The involvement of the trifluoperazine-binding site. , 1993, The Journal of biological chemistry.

[30]  Clara Franzini-Armstrong,et al.  STUDIES OF THE TRIAD I. Structure of the Junction in Frog Twitch Fibers , 1970 .