Abstract
The bacterial expression of human progastrin6–80 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 progastrin1–80 and to compare its biological activity with that of progastrin6–80 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 progastrin1–80 was expressed in Escherichia coli, collected on glutathione-agarose beads, and cleaved with enterokinase. Progastrin1–80 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 progastrin1–80 and progastrin6–80 in proliferation and migration assays with the mouse gastric cell line IMGE-5. We conclude that residues 1–5 of progastrin1–80 are not essential for biological activity.
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McQueen, K., Kovac, S., Ho, PK. et al. Preparation of Biologically Active Recombinant Human Progastrin1–80. J Protein Chem 21, 465–471 (2002). https://doi.org/10.1023/A:1021399003934
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DOI: https://doi.org/10.1023/A:1021399003934