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Research article

Identification of a novel 45 kDa protein (JP-45) from rabbit sarcoplasmic-reticulum junctional-face membrane

Francesco ZORZATO, Ayuk A. ANDERSON, Kai OHLENDIECK, Gabriele FROEMMING, Remo GUERRINI, Susan TREVES
Biochemical Journal Oct 15, 2000, 351 (2) 537-543; DOI: 10.1042/bj3510537
Francesco ZORZATO
Experimental and Diagnostic Medicine, General Pathology section, University of Ferrara, Via Borsari 46, 44100 Ferrara, ItalyDepartments of Anesthesia and Research, Basel Kantonsspital, Hebelstrasse 20, 4031 Basel, Switzerland
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Ayuk A. ANDERSON
Departments of Anesthesia and Research, Basel Kantonsspital, Hebelstrasse 20, 4031 Basel, Switzerland
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Kai OHLENDIECK
Department of Pharmacology, National University of Ireland, University College Dublin, Belfield, Dublin 4, Ireland
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Gabriele FROEMMING
Department of Pharmacology, National University of Ireland, University College Dublin, Belfield, Dublin 4, Ireland
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Remo GUERRINI
Department of Pharmaceutical Sciences and Biotechnology Center, Via Fossato di Mortara 17, University of Ferrara, 44100 Ferrara, Italy
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Susan TREVES
Departments of Anesthesia and Research, Basel Kantonsspital, Hebelstrasse 20, 4031 Basel, Switzerland
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  • For correspondence: susan.treves@unibas.ch
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Abstract

Using a biochemical/immunological approach to analyse the protein constituents of skeletal-muscle junctional-face membrane (JFM), we identified a 45kDa protein. Its N-terminal amino acid was blocked, but the amino acid sequence obtained from several peptides after proteolytic treatment did not significantly match that of any protein present in the SwissProt and NCBI (National Center for Biotechnology Information) databases. We synthesized a peptide whose sequence matched that of one of the peptides obtained after CNBr cleavage of the 45kDa protein; the peptide was conjugated to a carrier and used to raise antibodies. The antiserum was used to study in more detail the biochemical characteristics of the novel 45kDa protein. Analysis of the proteins present in different subcellular membrane fractions show that the novel 45kDa polypeptide: (i) is an integral membrane constituent present both in neonatal and adult skeletal-muscle sarcoplasmic reticulum; (ii) is selectively localized in the JFM; (iii) is not present in microsomes obtained from rabbit heart, liver or kidney. Immunoprecitation with anti-(45kDa protein) antibody indicates that the 45kDa protein is part of a complex which can be phosphorylated in vitro by the catalytic subunit of protein kinase A.

  • phosphorylated protein
  • skeletal muscle
  • transmembrane
  • The Biochemical Society, London © 2000
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October 2000

Volume: 351 Issue: 2

Biochemical Journal: 351 (2)
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Identification of a novel 45 kDa protein (JP-45) from rabbit sarcoplasmic-reticulum junctional-face membrane
Francesco ZORZATO, Ayuk A. ANDERSON, Kai OHLENDIECK, Gabriele FROEMMING, Remo GUERRINI, Susan TREVES
Biochemical Journal Oct 2000, 351 (2) 537-543; DOI: 10.1042/bj3510537
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Identification of a novel 45 kDa protein (JP-45) from rabbit sarcoplasmic-reticulum junctional-face membrane
Francesco ZORZATO, Ayuk A. ANDERSON, Kai OHLENDIECK, Gabriele FROEMMING, Remo GUERRINI, Susan TREVES
Biochemical Journal Oct 2000, 351 (2) 537-543; DOI: 10.1042/bj3510537

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Keywords

phosphorylated protein
skeletal muscle
transmembrane

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