Purification and structural characterization of progastrin-derived peptides from a human gastrinoma.
Several peptides derived from the gastrin-predicted preprohormone sequence were isolated from a human gastrinoma by gel permeation, anion exchange, and reverse phase chromatography. The peptides were identified and characterized structurally by a combination of radioimmunoassays, mass spectral analysis, and microsequence analysis. The largest peptide, progastrin-(1-35) (cryptagastrin), extends from the putative processing site for the signal peptidase to the double basic residues adjacent to the amino terminus of gastrin 34. A shorter form of this peptide, progastrin-(6-35) (cryptagastrin-(6-35), was also isolated in smaller amounts. In addition, sulfated and nonsulfated gastrin 17 amides (progastrin-(55-71)) and the glycine-extended nonsulfated gastrin 17 (progastrin-(55-72)) were identified by radioimmunoassay, and their structures were confirmed by mass spectral analysis. Isolation of cryptagastrin indicates that the signal peptide of human preprogastrin contains 21 amino acid residues, and progastrin, therefore, contains 80 amino acids. There is minimal processing of the cryptic peptide preceding the sequence of gastrin 34. An amidated gastrin form larger than gastrin 34 could contain 71 amino acids. No evidence was obtained for processing that would produce gastrins containing more than 34 but less than 71 amino acid residues.
Huebner
V D
VD
Beckman Research Institute of the City of Hope, Duarte, California 91010.
Jiang
R L
RL
Lee
T D
TD
Legesse
K
K
Walsh
J H
JH
Shively
J E
JE
Chew
P
P
Azumi
T
T
Reeve
J R
JR
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D010450:Endopeptidases / Q000378:metabolism; D015408:Gastrinoma / Q000737:chemistry*; D005755:Gastrins / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011863:Radioimmunoassay; D012697:Serine Endopeptidases; D016339:Spectrometry, Mass, Fast Atom Bombardment
D000596:Amino Acids; D005755:Gastrins; D008565:Membrane Proteins; D010455:Peptides; D011498:Protein Precursors; C013706:big gastrin; D010450:Endopeptidases; D012697:Serine Endopeptidases; C026515:type I signal peptidase