title: Isolation of endopeptidase-24.11 (EC 3.4.24.11, "enkephalinase") from the pig stomach. Hydrolysis of substance P, gastrin-releasing peptide 10, [Leu5] enkephalin, and [Met5] enkephalin.
abstract: The purpose of this investigation was to isolate the cell-surface enzyme endopeptidase-24.11 from the stomach wall of the pig and to examine the hydrolysis of the gastric neuropeptides. Endopeptidase-24.11 was isolated from gastric membranes by immunoadsorbent chromatography using a monoclonal antibody to porcine kidney endopeptidase-24.11. The enzyme was purified with a yield of 1.2 micrograms/g wet wt of fundic muscle. A single polypeptide chain of apparent subunit molecular weight of 90,000 was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gastric endopeptidase-24.11 hydrolyzed substance P, gastrin-releasing peptide 10, [Leu5] enkephalin, and [Met5] enkephalin by cleavage of peptide bonds on the N-terminal side of hydrophobic amino acids. The enzymatic activity was inhibited completely by phosphoramidon (10(-6) M) and strongly by 1,10-phenanthroline (10(-3) M), but was unaffected by captopril (10(-5) M).
authors: University of Washington, Department of Surgery, Seattle.
authors: N W
authors: NW
authors: Bunnett
authors: A J
authors: AJ
authors: Turner
authors: J
authors: J
authors: Hryszko
authors: R
authors: R
authors: Kobayashi
authors: J H
authors: JH
authors: Walsh
mesh_terms: D000818:Animals; D001839:Bombesin / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004743:Enkephalin, Leucine / Q000378:metabolism*; D004744:Enkephalin, Methionine / Q000378:metabolism*; D005753:Gastric Mucosa / Q000201:enzymology*; D008666:Metalloendopeptidases / Q000302:isolation & purification* / Q000502:physiology; D015260:Neprilysin; D010446:Peptide Fragments / Q000378:metabolism*; D010449:Peptide Mapping; D013373:Substance P / Q000378:metabolism*
chemical_list: D010446:Peptide Fragments; D013373:Substance P; D004744:Enkephalin, Methionine; D004743:Enkephalin, Leucine; C040794:neuromedin C; D008666:Metalloendopeptidases; D015260:Neprilysin; D001839:Bombesin