title: A novel proteinase, glucagon-degrading enzyme, secreted by a human pancreatic cancer cell line, HPC-YO.
abstract: Sixty-four kinds of cell lines were examined as to their ability to degrade glucagon using conditioned-media obtained from their protein-free cultures. Two human tumor cell lines were shown to produce this activity, and the cell line, HPC-YO, established from a human pancreatic carcinoma was shown to produce the highest level of activity. The glucagon-degrading enzyme (GDE) was purified from HPC-YO conditioned-medium by a combination of ion-exchange, gel filtration, and hydroxylapatite column chromatographies. The purified GDE also degraded vasoactive intestinal polypeptide (VIP) and secretin, however, it did not cleave EGF, gastrin, insulin, somatostatin, substance P, neurotensin, or growth hormone. The molecular weight of GDE is 83,000, as determined on SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of GDE was blocked, and the five partial amino acid sequences obtained on lysyl-endopeptidase digestion were determined to be N-L-T-E-E-Y-D-V-S-D-G-E-I-E-L-L-Y-E-K, V-E-T-Y-Y-D-L-L-F-E-K, L-Y-W-F-L-D-E-A-K, S-N-S-T-S-Y-V-K, and Y-Y-A-S-T-S-Y-D-D-T-Y-K. The same or homologous amino acid sequences have not been found in known proteins, demonstrating that GDE is a novel peptidase that degrades the secretin family: glucagon, VIP, and secretin.
authors.affiliation: Department of Cell Biology, Kyoto Prefectural University of Medicine.
authors.forename: N
authors.initials: N
authors.lastname: Yamaguchi
authors.forename: K
authors.initials: K
authors.lastname: Koyama
authors.forename: Y
authors.initials: Y
authors.lastname: Yamamura
authors.forename: A
authors.initials: A
authors.lastname: Okui
authors.forename: E
authors.initials: E
authors.lastname: Konishi
authors.forename: T
authors.initials: T
authors.lastname: Katayama
authors.forename: N
authors.initials: N
authors.lastname: Katsuragi
authors.forename: T
authors.initials: T
authors.lastname: Kurihara
authors.forename: H
authors.initials: H
authors.lastname: Nakazato
authors.forename: J
authors.initials: J
authors.lastname: Imanishi
mesh_terms: D000595:Amino Acid Sequence; D010450:Endopeptidases / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D010190:Pancreatic Neoplasms / Q000201:enzymology*; D012633:Secretin / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism*
chemical_list: D012633:Secretin; D014660:Vasoactive Intestinal Peptide; D010450:Endopeptidases; C093907:glucagon-degrading enzyme