Binding of pepsinogen to the 78 kDa gastrin binding protein.
An endogenous ligand of the 78 kDa gastrin-binding protein (GBP) has been purified from detergent extracts of porcine gastric mucosal membranes by ion exchange chromatography and preparative gel electrophoresis. The ligand bound to the GBP with high affinity (mean IC50 value of 0.31+/-0.09 microgram/ml, or 8 nM), as assessed by inhibition of cross-linking of iodinated gastrin2,17 to the GBP. Both the N- and C-terminal halves of the GBP, which had been expressed individually as glutathione-S-transferase fusion proteins in Escherichia coli, and purified on glutathione-agarose beads, bound the ligand. Two peptides derived from the ligand were purified by reversed-phase high-performance liquid chromatography (HPLC), and characterised by mass spectrometry and Edman sequencing. The peptides were 97% and 100% identical, respectively, to amino acids 119-157 and 199-219 of porcine pepsinogen A. Commercial samples of pepsinogen also bound to the GBP, with a mean IC50 value of 3.9+/-1. 2 micrograms/ml (100 nM). We conclude that the ligand is closely related, but not identical, to pepsinogen A.
Rorison
K A
KA
University Department of Surgery, Austin Campus, A & RMC, Studley Road, Heidelberg, Melbourne, Victoria 3084, Australia.
Neumann
G M
GM
Baldwin
G S
GS
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005753:Gastric Mucosa / Q000378:metabolism; D008024:Ligands; D063991:Mitochondrial Trifunctional Protein; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D020116:Pepsinogen A / Q000737:chemistry / Q000378:metabolism; D010435:Pepsinogens / Q000737:chemistry / Q000378:metabolism*; D010446:Peptide Fragments / Q000737:chemistry; D013552:Swine; D014357:Trypsin
D002352:Carrier Proteins; D008024:Ligands; D009097:Multienzyme Complexes; D010435:Pepsinogens; D010446:Peptide Fragments; D020116:Pepsinogen A; D063991:Mitochondrial Trifunctional Protein; D014357:Trypsin