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

Cite this

@article{bdf2c37416024362ab386bda5f48dde2,
title = "N-terminal domain of Bothrops asper Myotoxin II Enhances the Activity of Endothelin Converting Enzyme-1 and Neprilysin",
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.",
author = "Smith, {A Ian} and Rajapakse, {Niwanthi W} and Oded Kleifeld and Bruno Lomonte and Sikanyika, {Nkumbu L} and Spicer, {Alex J} and Hodgson, {Wayne C} and Conroy, {Paul J} and Small, {David H} and Kaye, {David M} and Parkington, {Helena C} and Whisstock, {James C} and Kuruppu, {Sanjaya Monath Sanjaya}",
year = "2016",
month = "3",
day = "2",
doi = "10.1038/srep22413",
language = "English",
volume = "6",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Publishing Group",

}

N-terminal domain of Bothrops asper Myotoxin II Enhances the Activity of Endothelin Converting Enzyme-1 and Neprilysin. / Smith, A Ian; Rajapakse, Niwanthi W; Kleifeld, Oded; Lomonte, Bruno; Sikanyika, Nkumbu L; Spicer, Alex J; Hodgson, Wayne C; Conroy, Paul J; Small, David H; Kaye, David M; Parkington, Helena C; Whisstock, James C; Kuruppu, Sanjaya Monath Sanjaya.

In: Scientific Reports, Vol. 6, 22413, 02.03.2016.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - N-terminal domain of Bothrops asper Myotoxin II Enhances the Activity of Endothelin Converting Enzyme-1 and Neprilysin

AU - Smith, A Ian

AU - Rajapakse, Niwanthi W

AU - Kleifeld, Oded

AU - Lomonte, Bruno

AU - Sikanyika, Nkumbu L

AU - Spicer, Alex J

AU - Hodgson, Wayne C

AU - Conroy, Paul J

AU - Small, David H

AU - Kaye, David M

AU - Parkington, Helena C

AU - Whisstock, James C

AU - Kuruppu, Sanjaya Monath Sanjaya

PY - 2016/3/2

Y1 - 2016/3/2

N2 - 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.

AB - 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.

UR - http://www.ncbi.nlm.nih.gov/pubmed/26931059

U2 - 10.1038/srep22413

DO - 10.1038/srep22413

M3 - Article

VL - 6

JO - Scientific Reports

JF - Scientific Reports

SN - 2045-2322

M1 - 22413

ER -