Preparation and Characterization of Recombinant Dolphin Fish (Coryphaena hippurus) Growth Hormone

A Paduel; N Chapnik-Cohen; A Gertler; Abigail Elizur

doi:10.1006/prep.1999.1088

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Preparation and Characterization of Recombinant Dolphin Fish (Coryphaena hippurus) Growth Hormone

Journal article

Peer reviewed

Preparation and Characterization of Recombinant Dolphin Fish (Coryphaena hippurus) Growth Hormone

A Paduel, N Chapnik-Cohen, A Gertler and Abigail Elizur

Protein Expression and Purification, Vol.16(3), pp.417-423

1999

DOI: https://doi.org/10.1006/prep.1999.1088

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Abstract

Biochemistry and Cell Biology

growth hormone

recombinant

dolphin fish

Coryphaena hippurus

Dolphin fish (Coryphaena hippurus) growth hormone (dfGH) cDNA encoding the mature protein was cloned in a pET11a expression vector and expressed in Escherichia coli BL21 cells upon induction with isopropyl-1-thio-β--galactopyranoside as an insoluble protein. The expressed protein, contained within the inclusion-body pellet, was solubilized in 4.5 M urea, refolded at pH 11.3 in the presence of catalytic amounts of cysteine, and purified to homogeneity, as evidenced by SDS-PAGE. Gel filtration on a Superdex column under nondenaturing conditions and amino-terminal analysis showed the purified protein to be monomeric methionyl-dfGH. Binding assays of the 125I-labeled dfGH to dolphin fish liver microsomal fraction resulted in high specific binding characterized by a Ka of 0.77 nM-1 and a Bmax of 285 fmol/mg microsomal fraction protein. The purified dfGH was capable of stimulating proliferation of FDC-P1-B9 cells transfected with rabbit growth hormone (GH) receptor. The maximal effect of dfGH was identical to that of human GH but their respective EC50 values were 28 nM versus 0.095 nM.

Details

Title: Preparation and Characterization of Recombinant Dolphin Fish (Coryphaena hippurus) Growth Hormone
Authors: A Paduel (Author) - Israel Oceanographic and Limnological Research
N Chapnik-Cohen (Author) - Hebrew University of Jerusalem
A Gertler (Author) - Hebrew University of Jerusalem
Abigail Elizur (Author) - University of the Sunshine Coast - Faculty of Science, Health and Education
Publication details: Protein Expression and Purification, Vol.16(3), pp.417-423
Publisher: Academic Press
Date published: 1999
DOI: 10.1006/prep.1999.1088
ISSN: 1046-5928
Organisation Unit: University of the Sunshine Coast, Queensland; Centre for Bioinnovation
Language: English
Record Identifier: 99449160202621
Output Type: Journal article

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Collaboration types: Domestic collaboration
Web Of Science research areas: Biochemical Research Methods; Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology

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