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Phosphorylation of the triadin cytoplasmic domain by CaM protein kinase in rabbit fast-twitch muscle sarcoplasmic reticulum

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Abstract

Skeletal muscle triadin is a sarcoplasmic reticulum (SR) membrane protein that had been shown to interact structurally and functionally at the cytoplasmic domain (amino acid residues 1–47) with the ryanodine receptor (RyR1), and to undergo phosphorylation by endogenous calmodulin protein kinase (CaM K II) in isolated terminal cisternae from rabbit fast-twitch muscle. Here we show that triadin cytoplasmic domain expressed as glutathione-S-transferase fusion protein, is a substrate of the protein kinase. This finding is corroborated by identification of a specific consensus sequence in the deduced amino sequence between residue 34 and 37 of triadin. Confirming the regulatory features of CaM K II, we show the phosphorylation of triadin cytoplasmic segment by the kinase, when converted to the autonomous form. We propose that triadin modulates RyR1 in a phosphorylation-dependent manner.

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References

  1. Knudson CM, Stang KK, Jorgensen AO, Campbell KP: Biochemical characterization and ultrastructural localization of a major junctional sarcoplasmic reticulum glycoprotein (triadin). J Biol Chem 268: 12637–12645, 1993

    Google Scholar 

  2. Kim CK, Caswell AH, Talvenehimo JA, Brandt NR: Isolation of a terminal cisternae protein which may link the dihyropyridine receptor to the junctional foot protein in skeletal muscle. Biochemistry 29: 9281–9289, 1980

    Google Scholar 

  3. Liu G, Pessah IN: Molecular interaction between ryanodine receptor and glycoprotein triadin involves redox cycling of functionally important hyperreactive sulfhydryls. J Biol Chem 269: 33028–33034, 1994

    Google Scholar 

  4. Okhura M, Furukawa K-I, Fujimori H, Kuruma A, Kawano S, Hiraoka M, Kuniyasu A, Nakayama H, Ohizumi Y: Dual regulation of the skeletal muscle ryanodine receptor by triadin and calsequestrin. Biochemistry 37: 12987–12993, 1998

    Google Scholar 

  5. Kuniyasu A, Kawano S, Hirayama Y, Ji Y-H, Xu K, Okhura M, Furukawa K-I, Ohizumi Y, Hiraoka M, Nakayama H: A new scorpion toxin (BmK-PL) stimulates Ca2+-release channel activity of skeletal muscle ryanodine receptor by an indirect mechanism. Biochem J 339: 343–350, 1999

    Google Scholar 

  6. Groh S, Marty I, Ottolia M, Prestipino G, Chapel A, Villaz M, Ronjat M: Functional interaction of the cytoplasmic domain of triadin with the skeletal ryanodine receptor. J Biol Chem 274: 12278–12283, 1999

    Google Scholar 

  7. Sacchetto R, Turcato F, Damiani E, Margreth A: Interaction of triadin with histidine-rich Ca2+-binding protein at the triadic junction in skeletal muscle fibers. J Muscle Res Cell Motil 20: 403–415, 1999

    Google Scholar 

  8. Hofmann S, Goldstein J, Orth K, Moomaw C, Slaughter C, Brown M: Molecular cloning of a histidine-rich Ca2+-binding protein of sarcoplasmic reticulum that contains highly conserved repeated elements. J Biol Chem 264: 18083–18090, 1989

    Google Scholar 

  9. Simmerman H, Jones L: Phospholamban: Protein structure, mechanism of action and role in cardiac function. Physiol Rev 78: 921–947, 1998

    Google Scholar 

  10. Damiani E, Picello E, Saggin L, Margreth A: Identification of triadin and of histidine-rich Ca2+-binding protein as substrates of 60 kDa calmodulin-dependent protein kinase in junctional terminal cisternae of sarcoplasmic reticulum of rabbit fast muscle. Biochem Biophys Res Commun 209: 457–465, 1995

    Google Scholar 

  11. Braun A, Schulman H: The multifunctional calcium/calmodulin dependent protein kinase: From form to function. Annu Rev Physiol 57: 417–444, 1995

    Google Scholar 

  12. Knudson C, Stang K, Moomaw C, Slaughter C, Campbell KP: Primary structure and topological analysis of a skeletal muscle-specific junctional sarcoplasmic reticulum glycoprotein (Triadin). J Biol Chem 268: 12646–12654, 1993

    Google Scholar 

  13. Guo W, Campbell KP: Association of triadin with the ryanodine receptor and calsequestrin in the lumen of the sarcoplasmic reticulum. J Biol Chem 270: 9027–9030, 1995

    Google Scholar 

  14. Saito A, Seiler S, Chu A, Fleischer S: Preparation and morphology of sarcoplasmic reticulum terminal cisternae from rabbit skeletal muscle. J Cell Biol 99: 875–885, 1984

    Google Scholar 

  15. Damiani E, Volpe P, Margreth A: Coexpression of two isoforms of calsequestrin in rabbit slow-twitch muscle. J Muscle Res Cell Motil 11: 522–530, 1990

    Google Scholar 

  16. Tuana B, MacLennan DH: Isolation of the calmodulin dependent protein kinase system from rabbit skeletal muscle sarcoplasmic reticulum. FEBS Lett 235: 219–223, 1988

    Google Scholar 

  17. Damiani E, Sacchetto R, Margreth A: Phosphorylation of anchoring protein by calmodulin protein kinase associated to the sarcoplasmic reticulum of rabbit fast-twitch muscle. Biochem Biophys Res Commun 279: 181–189, 2000

    Google Scholar 

  18. Campbell N, Sargent JR: Estimation of total protein by the method of Lowry et al. (1951). In: P.N. Campbell, J.R. Sargent (eds). Techniques in Protein Biosynthesis. Academic Press, London, 1967, pp 299–311

    Google Scholar 

  19. Damiani E, Tobaldin G, Bortoloso E, Margreth A: Functional behaviour of the ryanodine receptor/Ca2+-release channel in vesiculated derivatives of the junctional membrane of terminal cisternae of rabbit fast muscle sarcoplasmic reticulum. Cell Calcium 22: 129–151, 1997

    Google Scholar 

  20. Damiani E, Margreth A: Pharmacological clues to calmodulin-mediated activation of skeletal ryanodine receptor using [3H]-ryanodine binding. J Muscle Res Cell Motil 21: 1–8, 2000

    Google Scholar 

  21. Laemmli U-K: Cleavage of structural protein during the assembly of the head of bacteriophage. Nature 277: 680–685, 1970

    Google Scholar 

  22. Sacchetto R, Margreth A, Pelosi M, Carafoli E: Colocalization of the dihydropyridine receptor, the plasma-membrane calcium ATPase isoform 1 and the sodium/calcium exchanger to the junctional-membrane domain of transverse tubules of rabbit skeletal muscle. Eur J Biochem 237: 483–488, 1996

    Google Scholar 

  23. Bayer K-U, Harbers K, Schulman H: αKAP is an anchoring protein for a novel CaM kinase II isoform in skeletal muscle. EMBO J 19: 5598–5605, 1998

    Google Scholar 

  24. Kim DH, Sreter FA, Ikemoto N: Involvement of 60 kDa phosphoprotein in the regulation of Ca2+-release from sarcoplasmic reticulum of normal and malignant hyperthermia susceptible pig muscles. Biochim Biophys Acta 945: 246–252, 1988

    Google Scholar 

  25. Takasago T, Imagawa T, Furukawa K-I, Ogurusu T, Shigekawa M: Regulation of the cardiac ryanodine receptor by protein-kinase dependent phosphorylation. J Biochem 109: 163–170, 1991

    Google Scholar 

  26. Witcher D, Kovacs R, Schulman H, Cefali D, Jones LR: Unique phosphorylation site on the cardiac ryanodine receptor regulates calcium channel activity. J Biol Chem 266: 11144–11152, 1991

    Google Scholar 

  27. Strand M, Louis C, Mickelson J: Phosphorylation of the porcine skeletal and cardiac muscle sarcoplasmic reticulum ryanodine receptor. Biochim Biophys Acta 1175: 319–326, 1993

    Google Scholar 

  28. Leddy JJ, Murphy BJ, Yi Q, Doucet J-P, Pratt C, Tuana B: A 60 kDa polypeptide of skeletal muscle sarcoplasmic reticulum is a calmodulin-dependent protein kinase that associates with and phosphorylates several membrane proteins. Biochem J 295: 849–856, 1993

    Google Scholar 

  29. Dzhura I, Wu Y, Colbran R, Balser J, Anderson M: Calmodulin kinase determines calcium dependent facilitation of L-type calcium channels. Nature Cell Biology 2: 173–177, 2000

    Google Scholar 

  30. Berchtold MW, Brinkmeier H, Muntener M: Calcium ion in skeletal muscle: Its crucial role for muscle function, plasticity and disease. Physiol Rev 80: 1215–1265, 2000

    Google Scholar 

  31. Hain J, Nath S, Mayrleitner M, Fleischer S: Phosphorylation modulates the function of the calcium release channel of sarcoplasmic reticulum from skeletal muscle. Biophys J 67: 1823–1833, 1994

    Google Scholar 

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Authors and Affiliations

  1. NRC Unit for Muscle Biology and Physiopathology, Department of Experimental Biomedical Sciences, University of Padova, viale G. Colombo 3, 35121, Padova, Italy

    Pierangelo Colpo, Alessandra Nori, Roberta Sacchetto, Ernesto Damiani & Alfredo Margreth

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  1. Pierangelo Colpo
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  2. Alessandra Nori
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  3. Roberta Sacchetto
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  4. Ernesto Damiani
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  5. Alfredo Margreth
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Correspondence to Alfredo Margreth.

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Colpo, P., Nori, A., Sacchetto, R. et al. Phosphorylation of the triadin cytoplasmic domain by CaM protein kinase in rabbit fast-twitch muscle sarcoplasmic reticulum. Mol Cell Biochem 223, 139–145 (2001). https://doi.org/10.1023/A:1017987015807

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