Positional effects in the neprilysin (neutral endopeptidase) reaction

Tracey Quay, Clive A. Slaughter, Thomas P. Davis, Bradley J. Merrill, Louis B. Hersh

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Neprilysin is a peptidase which has a specificity directed toward cleavage on the amino side of hydrophobic residues. In addition an active site arginine on the enzyme can interact with the C-terminal carboxylate of a substrate. The importance of the position of the hydrophobic residue relative to the C-terminus of the substrate has been investigated using a series of peptides containing one or two cleavage sites. With a hexapeptide series succ-(Gly)x-Phe-(Gly)y-OH, where x = 1 to 5 and y = 5 to 1 respectively, a ∼25-fold increase in the specificity constant kcat/Km was observed when Phe was adjacent to the C-terminal Gly residue. With peptide-free acids containing two cleavable bonds (X and Y) of the type succ-Gly-X-Gly-Y-Gly-OH, cleavage was observed at the Y residue. However, when the two cleavable bonds were adjacent, succ-Gly-Gly-X-Y-Gly-OH, cleavage of a tripeptide was observed even when the residue in position X was one cleaved poorly when presented as the sole cleavage site. These results demonstrate a preference by the enzyme for the placement of a hydrophobic residue in the P′2 position.

Original languageEnglish (US)
Pages (from-to)133-136
Number of pages4
JournalArchives of Biochemistry and Biophysics
Volume308
Issue number1
DOIs
StatePublished - Jan 1 1994
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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