Cloning and characterization of a new isoform of skeletal muscle triadin.
We have shown that several isoforms of triadin, a protein involved in calcium release process through the ryanodine receptor, are expressed in rat skeletal muscle, and we have cloned two of these isoforms. One is the rat homolog of the 95-kDa triadin identified in rabbit skeletal muscle, and the second one, shorter, is a truncated form of the previous one, but with a new unique COOH-terminal end. We propose to name the two proteins identified here Trisk 95 and Trisk 51. We have produced antibodies specific to each isoform. Using these antibodies, we have shown that the newly identified protein, Trisk 51, is actually expressed in adult rat skeletal muscle and also in rat embryo skeletal muscle. Immunofluorescent labeling of rat skeletal muscle with anti-Trisk 95, anti-Trisk 51, or anti-ryanodine receptor antibodies shows a similar localization of these proteins, in the tissue. Transfection of L6 cells with cDNA of Trisk 51 or Trisk 95 leads to the expression of proteins with the expected molecular weight, identical to those detected in rat skeletal muscle. Both proteins appear during differentiation of satellite cells in myotubes which may indicate the involvement of these two isoforms in the building of a functional calcium release machinery.
Marty
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I
Departement de Biologie Moléculaire et Structurale/Canaux Ioniques et Signalisation, INSERM E9931, Commissariat à l'Energie Atomique, Grenoble, 17 rue des Martyrs, Grenoble 38054, France. imarty@cea.fr
Thevenon
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D
Scotto
C
C
Groh
S
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Sainnier
S
S
Robert
M
M
Grunwald
D
D
Villaz
M
M
D000595:Amino Acid Sequence; D000818:Animals; D002352:Carrier Proteins; D002451:Cell Compartmentation; D002454:Cell Differentiation; D002458:Cell Fractionation; D005455:Fluorescent Antibody Technique; D015723:Gene Library; D006031:Glycosylation; D047908:Intracellular Signaling Peptides and Proteins; D008861:Microsomes / Q000737:chemistry; D008969:Molecular Sequence Data; D009124:Muscle Proteins / Q000235:genetics / Q000302:isolation & purification*; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000166:cytology / Q000196:embryology; D020033:Protein Isoforms / Q000235:genetics / Q000302:isolation & purification; D011499:Protein Processing, Post-Translational; D051381:Rats; D019837:Ryanodine Receptor Calcium Release Channel / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D002352:Carrier Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009124:Muscle Proteins; D020033:Protein Isoforms; D019837:Ryanodine Receptor Calcium Release Channel; C498271:Trdn protein, rat; C070048:triadin