C-terminal fragments of the gastrin-releasing peptide precursor stimulate cell proliferation via a novel receptor.
There are many precedents for the production from a single precursor of multiple peptides, with independent receptors and different bioactivities. Gastrin-releasing peptide (GRP) is initially synthesized as amino acids 1-27 of a 125-residue precursor, proGRP, and is subsequently cleaved and amidated to form GRP18-27. We investigated the hypothesis that C-terminal proGRP peptides are also biologically active. Human proGRP18-125 was expressed in Escherichia coli as a glutathione S-transferase fusion protein. Recombinant proGRP18-125 stimulated proliferation and migration of the human colorectal carcinoma cell line DLD-1. The observations that an antagonist selective for the GRP receptor did not inhibit activity in either proliferation or migration assays and that the recombinant peptide did not bind to either the GRP receptor or orphan receptor BRS-3 indicated that neither activity was mediated by the known GRP receptors. Recombinant human proGRP31-125 and proGRP42-98 were also prepared and shown to stimulate proliferation of DLD-1 cells and the human prostate carcinoma cell line DU145. The synthetic peptides proGRP47-68 and [Tyr79]proGRP80-97 stimulated inositol phosphate production, MAPK kinase activity, and proliferation and migration of DLD-1 cells. Binding sites for both radioiodinated synthetic peptides were demonstrated on DLD-1 cells. Each peptide was able to compete with the other for binding, and a GRP receptor antagonist did not inhibit binding of either peptide. We conclude that peptides derived from the C terminus of proGRP are biologically active and that their activity is mediated by a receptor distinct from the two known GRP receptors.
Patel
Oneel
O
Department of Surgery, Austin Health, Studley Road, Heidelberg, Victoria 3084, Australia. grahamsb@unimelb.edu.au
Dumesny
Chelsea
C
Shulkes
Arthur
A
Baldwin
Graham S
GS
D001665:Binding Sites; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D017124:Conserved Sequence; D006801:Humans; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology*; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D011498:Protein Precursors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology*; D018004:Receptors, Bombesin / Q000378:metabolism*; D011994:Recombinant Proteins / Q000138:chemical synthesis / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects
D010446:Peptide Fragments; D010455:Peptides; D011498:Protein Precursors; D018004:Receptors, Bombesin; D011994:Recombinant Proteins; C047897:gastrin-releasing peptide precursor