Fluorogenic substrates for the enkephalin-degrading neutral endopeptidase (Enkephalinase)

Robert S. Rush, Michael Mitas, James C. Powers, Takumi Tanaka, Louis B. Hersh

Research output: Contribution to journalArticlepeer-review

14 Scopus citations


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.

Original languageEnglish
Pages (from-to)390-399
Number of pages10
JournalArchives of Biochemistry and Biophysics
Issue number2
StatePublished - Jun 1984

Bibliographical note

Funding Information:
1 This research was supported in part by Grant DA 02243 from National Institute of Drug Abuse (L.B.H.); Grant I-391 from the Robert A. Welch Foundation, Houston, Texas (L.B.H.); and Grant HL 18679 from the National Institute of Health (J.C.P.). ‘To whom correspondence regarding this manuscript should be addressed.

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology


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