Purification and structural determination of urinary NH2-terminal big gastrin fragments.
We previously demonstrated that there existed extremely abundant NH2-terminal big gastrin immunoreactivity (NT G-34-IR) in human urine. This report describes the purification and sequence of NT G-34-IR from the urine of an achlorhydric patient. The purification was carried out by a combination of Sep-Pak C18 cartridges, Sephadex G-25, and HPLC steps using a radioimmunoassay specific for NH2-terminus of G-34 and ultraviolet absorption at 214 nm as monitors. Three peptides were isolated. The amino acid analysis, mass spectrometry, and sequence analysis confirmed the structures of urinary NT G-34 fragments being less than Glu-Leu-Gly-Pro-Gln-Gly-Pro-Pro, less than Glu-Leu-Gly-Pro-Gln-Gly- Pro-Pro-His, and less than Glu- Leu-Gly-Pro-Gln-Gly-Pro-Pro-His-Leu. NH2-terminal octapeptide of G-34 was the main component of urinary NT G-34-IR.
Higashimoto
Y
Y
Second Department of Internal Medicine, Osaka University Medical School, Japan.
Himeno
S
S
Shinomura
Y
Y
Nagao
K
K
Tamura
T
T
Tarui
S
S
D000126:Achlorhydria / Q000652:urine*; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D005755:Gastrins / Q000652:urine*; D006801:Humans; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000652:urine*; D011498:Protein Precursors / Q000652:urine*; D011863:Radioimmunoassay
D000596:Amino Acids; D005755:Gastrins; D010446:Peptide Fragments; D011498:Protein Precursors; C013706:big gastrin