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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References cited
Agellon, L.B. and Chen, T.T. 1986. Rainbow trout growth hormone: molecular cloning of cDNA expression inEscherichia coli DNA 5: 463–471.
Ball, J.N. 1965. Partial hypophysectomy in the teleostPoecilia: separate identities of teleostean growth hormone and teleostean prolactin-like hormone. Gen. Comp. Endocrinol. 5: 654–661.
Bolton, J.P., Takahashi, A., Kawauchi, H., Kubota, J. and Hirano, T. 1986. Development and validation of a salmon growth hormone radioimmunoassay. Gen. Comp. Endocrinol. 62: 230–238.
Breton, B., Prunet, P. and Reinaud, P. 1978. Sexual differences in salmon gonadotropin. Ann. Biol. Anim. Bioch. Biophys. 18: 759–765.
Cook, A.F., Wilson, S.W. and Peter, R.E. 1983. Development and validation of a carp growth hormone radioimmunoassay. Gen. Comp. Endocrinol. 50: 335–347.
Dubois, M.P. and Herlant, M. 1968. Caractères cytologiques des cellules gonadotropes, thyréotropes, corticotropes, somatotropes et des cellules à prolactine présentes dans le lobe antérieur de l'hypophyse des bovins. Ann. Biol. Anim. Bioch. Biophys. 8: 5–20.
Farbridge, K.J., Burke, M.G. and Leatherland, J.F. 1985. Seasonal changes in the structure of the adenohypophysis of the brown bullhead (Ictalurus nebulosus). Cytobios 44: 49–66.
Farmer, S.W., Papkoff, H., Hayashida, T., Bewley, T.A., Bern, H.A. and Li, C.H. 1976. Purification and properties of teleost growth hormone. Gen. Comp. Endocrinol. 30: 91–100.
Fryer, J.N. 1979. A radioreceptor assay for purified teleost growth hormone. Gen. Comp. Endocrinol. 39: 123–130.
Fryer, J.N. and Bern, H.A. 1979. Growth hormone binding to tissues of normal and stunted juvenile coho Salmon, (Oncorhynchus kisutch). J. Fish Biol. 15: 527–533.
Gabe, M. 1968. Techniques Histologiques. Masson et Cie, Paris.
Gerasimo, P., Djiane, J. and Kelly, P.A. 1979. Titration of total binding sites for growth hormone in rabbit liver. Quantitative modifications of these sites during pregnancy. Mol. Cell. Endocrinol. 13: 11–23.
Hirano, T., Prunet, P., Kawauchi, H., Takahashi, A., Ogasawara, T., Kubota, J. Nishioka, R.S. Bern, H.A., Takada, K. and Ishii, S. 1985. Development and validation of a Salmon prolactin radioimmunoassay. Gen. Comp. Endocrinol. 59: 266–276.
Kawauchi, H. Moriyama, S., Yasuda, A. Yamaguchi, K., Shirahata, K., Kubota, J. and Hirano, T. 1986. Isolation and characterization of chum salmon growth hormone. Arch. Biochem. Biophys. 244: 542–552.
Kelly, P.A., Leblanc, G. and Djiane, J. 1979. Estimation of total prolactin-binding sites afterin vitro desaturation. Endocrinology 104: 1631–1638.
Kishida, M. Hirano, T., Kubota, J., Hasegawa, S., Kawauchi, H., Yamaguchi, K. and Shirahata, K. 1987. Isolation of two forms of growth hormone secreted from eel pituitariesin vitro. Gen. Comp. Endocrinol. 65: 478–488.
Komourdjian, M.P. and Idler, D.R. 1979. Chum salmon pituitary fractions: somatotropic activity and cytoimmuno-fluorescence studies. Gen. Comp. Endocrinol. 37: 343–349.
Laemmli, U.K. 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, Lond. 227: 680–685.
Lowry, O.H., Rosebrough, N.J., Lewis Farr, A. and Randall, R.J. 1951. Protein measurement with the folin phenol reagent. J. Biol. Chem. 193: 265–275.
Nicoll, C.S., Steiny, S.S., King, D.S., Nishioka, R.S., Mayer, G.L., Eberhardt, N.L., Baxter, J.D., Yamanaka, M.K., Miller, J.A., Seilhamer, J.J., Schilling, J.W. and Johnson, L.K. 1987. The primary structure of coho salmon growth hormone and its cDNA. Gen. Comp. Endocrinol. 68: 387–399.
Olivereau, M. 1970. Stimulation des cellules somatotropes de l'hypophyse de la carpe après un jeune prolongé. C.R. Acad. Sci. Paris 270: 2343–2346.
Ouchterlony, O. 1967.In Handbook of Experimental Immunology, p. 655. Edited by Weir.
Specker, J.L., King, D.S., Nishioka, R.S., Shizahata, K., Yamaguchi, K. and Bern, H.A. 1985. Isolation and partial characterization of a pair of prolactins releasedin vitro by the pituitary of a cichlid fish,Oreochromis mossambicus. Proc. Nat. Acad. Sci. U.S.A. 82: 7490–7494.
Sumpter, J.P. and Donaldson, E.M. 1986. The development and validation of a radioimmunoassay to measure plasma ACTH levels in salmonid fishes. Gen. Comp. Endocrinol. 62: 367–376.
Tarpey, J.F. and Nicoll, C.S. 1985. Characterization of hepatic growth hormone binding sites in two fish species,Gillichthys mirabilis (Teleostei) andAcipenser transmontanus (Chondrostei). Gen. Comp. Endocrinol. 60: 39–50.
Wagner, G.F. and McKeown, B.A. 1983. The immuno-cytochemical localisation of pituitary somatotrops in the genusOncorhynchus using an antiserum to growth hormone of chum salmon (Oncorhynchus keta). Cell. Tiss. Res. 231: 693–697.
Wagner, G.F. Fargher, R.C., Brown, J.C. and McKeown, B.A. 1985. Further characterization of growth hormone from the chum salmon (Oncorhynchus keta). Gen. Comp. Endocrinol. 60: 27–34.
Wagner, G.F. and McKeown, B.A. 1986. Development of a salmon growth hormone radioimmunoassay. Gen. Comp. Endocrinol. 62: 452–458.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
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
Issue Date:
DOI: https://doi.org/10.1007/BF00004713