Allosteric inhibition of the neuropeptidase neurolysin

J Biol Chem. 2014 Dec 19;289(51):35605-19. doi: 10.1074/jbc.M114.620930. Epub 2014 Nov 5.


Neuropeptidases specialize in the hydrolysis of the small bioactive peptides that play a variety of signaling roles in the nervous and endocrine systems. One neuropeptidase, neurolysin, helps control the levels of the dopaminergic circuit modulator neurotensin and is a member of a fold group that includes the antihypertensive target angiotensin converting enzyme. We report the discovery of a potent inhibitor that, unexpectedly, binds away from the enzyme catalytic site. The location of the bound inhibitor suggests it disrupts activity by preventing a hinge-like motion associated with substrate binding and catalysis. In support of this model, the inhibition kinetics are mixed, with both noncompetitive and competitive components, and fluorescence polarization shows directly that the inhibitor reverses a substrate-associated conformational change. This new type of inhibition may have widespread utility in targeting neuropeptidases.

Keywords: Allosteric Regulation; Hydrolase; Metalloprotease; Neurochemistry; Neuropeptide; Peptidase; X-ray Crystallography.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Allosteric Regulation*
  • Allosteric Site
  • Animals
  • Binding Sites / genetics
  • Biocatalysis / drug effects
  • Catalytic Domain
  • Crystallography, X-Ray
  • Enzyme Inhibitors / chemical synthesis
  • Enzyme Inhibitors / chemistry*
  • Enzyme Inhibitors / pharmacology
  • Fluorescence Polarization
  • Kinetics
  • Metalloendopeptidases / chemistry*
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism
  • Models, Chemical
  • Models, Molecular
  • Molecular Structure
  • Mutation, Missense
  • Protein Binding
  • Protein Structure, Tertiary*
  • Rats
  • Substrate Specificity


  • Enzyme Inhibitors
  • Metalloendopeptidases
  • neurolysin

Associated data

  • PDB/1I1I
  • PDB/1R4L