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Purification of chinook salmon (Oncorhynchus tshawytscha) GH for receptor study

Fish Physiology and Biochemistry volume 7, pages 243–251 (1989)Cite this article

Abstract

A method for the purification of chinook Salmon (Oncorhynchus tshawytscha) GH, which retains its biological activity, is described. The biological activity was investigated with an established radioreceptor assay using liver membranes from pregnant rabbits and bovine GH as standard and labelled hormone. The enrichment of the preparation was checked with electrophoresis (SDS-PAGE). Extraction and further steps were carried out using low molarity alkaline buffer (pH 8–10, M = 100 mM). Three chromatography steps were performed (Concanavalin-A sepharose, Bio-gel P60, DEAE). Ion exchange chromatography was performed under isocratic conditions (using a 50 cm column). Two isoforms (sGH1 and sGH2) were isolated. The purification yield is 0.7% compared to lyophilized pituitaries. The molecule is homogeneous in SDS-PAGE. Contamination by prolactin, gonadotrophin and corticotrophin is negligible (< 0.5%). It could be demonstrated that the biological activity of the preparation is maintained since this preparation stimulates the growth of juvenile trout (Salmo gairdneri) and binds specifically (35%) to trout liver membranes.

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Affiliations

  1. Laboratoire de Physiologie des Poissons, INRA, Campus de Beaulieu, 35042, Rennes Cedex, France

    Pierre-Yves Le Bail & Geneviève Boulard

  2. Laboratoire de Physiologie Animale, INRA, 9 place Viala, 34060, Montpellier, France

    Bruno Barenton

  3. Station de Pathologie de la Reproduction, INRA, 37380, Nouzilly, France

    Michel Zygmunt

Authors
  1. Pierre-Yves Le Bail
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  2. Geneviève Boulard
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  3. Bruno Barenton
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  4. Michel Zygmunt
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Le Bail, PY., Boulard, G., Barenton, B. et al. Purification of chinook salmon (Oncorhynchus tshawytscha) GH for receptor study. Fish Physiol Biochem 7, 243–251 (1989). https://doi.org/10.1007/BF00004713

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  • DOI: https://doi.org/10.1007/BF00004713

Keywords

  • fish
  • salmon
  • growth hormone
  • biological activity
  • radioreceptor assay
  • purification techniques