N-terminal domain of Bothrops asper Myotoxin II Enhances the Activity of Endothelin Converting Enzyme-1 and Neprilysin

Sci Rep. 2016 Mar 2:6:22413. doi: 10.1038/srep22413.

Abstract

Neprilysin (NEP) and endothelin converting enzyme-1 (ECE-1) are two enzymes that degrade amyloid beta in the brain. Currently there are no molecules to stimulate the activity of these enzymes. Here we report, the discovery and characterisation of a peptide referred to as K49-P1-20, from the venom of Bothrops asper which directly enhances the activity of both ECE-1 and NEP. This is evidenced by a 2- and 5-fold increase in the Vmax of ECE-1 and NEP respectively. The K49-P1-20 concentration required to achieve 50% of maximal stimulation (AC50) of ECE-1 and NEP was 1.92 ± 0.07 and 1.33 ± 0.12 μM respectively. Using BLITZ biolayer interferometry we have shown that K49-P1-20 interacts directly with each enzyme. Intrinsic fluorescence of the enzymes change in the presence of K49-P1-20 suggesting a change in conformation. ECE-1 mediated reduction in the level of endogenous soluble amyloid beta 42 in cerebrospinal fluid is significantly higher in the presence of K49-P1-20 (31 ± 4% of initial) compared with enzyme alone (11 ± 5% of initial; N = 8, P = 0.005, unpaired t-test). K49-P1-20 could be an excellent research tool to study mechanism(s) of enzyme stimulation, and a potential novel drug lead in the fight against Alzheimer's disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alanine / metabolism
  • Amino Acid Sequence
  • Endothelin-Converting Enzymes / metabolism*
  • Enzyme Activation / drug effects
  • Enzyme Assays
  • Group II Phospholipases A2 / chemistry*
  • Group II Phospholipases A2 / pharmacology*
  • HEK293 Cells
  • Humans
  • Kinetics
  • Neprilysin / metabolism*
  • Peptides / chemistry
  • Peptides / metabolism
  • Protein Domains
  • Reptilian Proteins / chemistry*
  • Reptilian Proteins / pharmacology*
  • Structure-Activity Relationship

Substances

  • Peptides
  • Reptilian Proteins
  • Group II Phospholipases A2
  • myotoxin II, Bothrops asper
  • Neprilysin
  • Endothelin-Converting Enzymes
  • Alanine