The human gastrin precursor. Characterization of phosphorylated forms and fragments.
There is a potential phosphorylation site in the C-terminal region of the precursor for the acid-stimulating hormone gastrin, which is immediately adjacent to an important cleavage point. In the present study we have sought to identify, separate, quantify and characterize phosphorylated and unphosphorylated forms of human progastrin and its fragments. Identification was made by two radioimmunoassays: (a) a novel assay employing an antibody raised to intact human progastrin; and (b) an assay using antibody reacting with the C-terminal tryptic fragment of human progastrin, as well as progastrin itself. Two forms of human progastrin isolated from a gastrinoma were separated by ion-exchange h.p.l.c., and had similar elution positions on reverse-phase h.p.l.c. and on gel filtration. The more acidic peptide contained close to equimolar amounts of phosphate. On trypsinization, peptides were released that co-eluted on ion-exchange h.p.l.c. with, and had the immunochemical properties of, naturally occurring C-terminal fragments of progastrin. One of the latter was isolated and shown by Edman degradation after derivatization with ethanethiol to have the sequence Ser (P)-Ala-Glu-Asp-Glu-Asn. Similar peptides occur in antral mucosa resected from ulcer patients. The unphosphorylated forms of progastrin predominated, whereas the phosphorylated forms of the C-terminal fragments were predominant. This distribution could be explained by preferential cleavage of phosphorylated progastrin. We conclude that in human progastrin, Ser-96 can occur in the phosphorylated form; this residue immediately follows a pair of basic residues (Arg-Arg) that are cleaved during synthesis of the biologically active product.
Varro
A
A
Physiological Laboratory, University of Liverpool, U.K.
Desmond
H
H
Pauwels
S
S
Gregory
H
H
Young
J
J
Dockray
G J
GJ
D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D005753:Gastric Mucosa / Q000032:analysis; D015408:Gastrinoma / Q000032:analysis; D005755:Gastrins / Q000302:isolation & purification*; D006801:Humans; D008113:Liver Neoplasms / Q000032:analysis; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000302:isolation & purification; D010766:Phosphorylation; D011498:Protein Precursors / Q000302:isolation & purification*; D014357:Trypsin
D005755:Gastrins; D010446:Peptide Fragments; D011498:Protein Precursors; C013706:big gastrin; D014357:Trypsin