title: Fluorogenic substrates for the enkephalin-degrading neutral endopeptidase (Enkephalinase).
abstract: Rat brain neutral endopeptidase ("Enkephalinase") was shown to hydrolyze a series of fluorogenic substrates of the general structure 2-aminobenzoyl-(amino acid)n- leucylalanylglycine -4- nitrobenzylamide . The hydrolysis of these substrates was competitively inhibited by Leu5-enkephalin, demonstrating that these are indeed substrates for the rat brain neutral endopeptidase. Cleavage of the fluorogenic substrates yielded leucylalanylglycine -4- nitrobenzylamide as a common product. In addition, a series of inhibitors previously shown to inhibit thermolysin-like enzymes inhibited the hydrolysis of both Leu5-enkephalin and the synthetic substrates. The results of this study (a) demonstrate that the enkephalin-degrading endopeptidase is similar in specificity to thermolysin, (b) provide a continuous sensitive assay system for the enzyme, and (c) point out the potential use of this substrate class for probing the specificity of the enzyme.
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology; D010450:Endopeptidases / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000138:chemical synthesis; D006868:Hydrolysis; D015260:Neprilysin; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism*; D011480:Protease Inhibitors; D051381:Rats; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides; D011480:Protease Inhibitors; D010450:Endopeptidases; D015260:Neprilysin