Please use this identifier to cite or link to this item: http://ahro.austin.org.au/austinjspui/handle/1/9462
Title: Preparation of biologically active recombinant human progastrin(1-80).
Authors: McQueen, Kim;Kovac, Suzana;Ho, Po-Ki;Rorison, Kristy;Pannequin, Julie;Neumann, Greg;Shulkes, Arthur;Baldwin, Graham S
Affiliation: University of Melbourne Department of Surgery, Austin and Repatriation Medical Centre, Melbourne, Victoria, Australia.
Issue Date: 1-Oct-2002
Citation: Journal of Protein Chemistry; 21(7): 465-71
Abstract: The bacterial expression of human progastrin(6-80) has been reported previously [Baldwin, G.S. et al. (2001) J. Biol. Chem. 276: 7791-7796]. The aims of the present study were to prepare full-length recombinant human progastrin(1-80) and to compare its biological activity with that of progastrin(6-80) in vitro, to determine whether or not the N-terminal five amino acids contributed to activity. A fusion protein of glutathione-S-transferase and human progastrin(1-80) was expressed in Escherichia coli, collected on glutathione-agarose beads, and cleaved with enterokinase. Progastrin(1-80) was purified by reversed-phase and anion exchange HPLC and characterized by radioimmunoassay, amino acid sequencing, and mass spectrometry. No differences were detected in the extent of stimulation by progastrin(1-80) and progastrin(6-80) in proliferation and migration assays with the mouse gastric cell line IMGE-5. We conclude that residues 1-5 of progastrin(1-80) are not essential for biological activity.
Internal ID Number: 12523650
URI: http://ahro.austin.org.au/austinjspui/handle/1/9462
URL: http://www.ncbi.nlm.nih.gov/pubmed/12523650
Type: Journal Article
Subjects: Amino Acid Sequence
Amino Acids.analysis
Animals
Cell Division.drug effects
Cell Line, Transformed
Cell Movement.drug effects
Chromatography, High Pressure Liquid.methods
Escherichia coli.genetics.metabolism
Gastric Mucosa.cytology
Gastrins.biosynthesis.genetics.pharmacology
Glutathione Transferase.genetics
Humans
Mice
Mice, Transgenic
Molecular Sequence Data
Protein Precursors.biosynthesis.genetics.pharmacology
Radioimmunoassay
Recombinant Fusion Proteins.biosynthesis.genetics.pharmacology
Spectrometry, Mass, Electrospray Ionization
Appears in Collections:Journal articles

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