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

A Ian Smith, Niwanthi W Rajapakse, Oded Kleifeld, Bruno Lomonte, Nkumbu L Sikanyika, Alex J Spicer, Wayne C Hodgson, Paul J Conroy, David H Small, David M Kaye, Helena C Parkington, James C Whisstock, Sanjaya Monath Sanjaya Kuruppu

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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 muM 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.
Original languageEnglish
Article number22413
Number of pages10
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 2 Mar 2016

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