A Simple Method for Isolating Human Endothelin Converting Enzyme Free from Contamination by Neutral Endopeptidase 24.11

1995 ◽  
Vol 207 (1) ◽  
pp. 355-362 ◽  
Author(s):  
R. Corder ◽  
N. Khan ◽  
V.J. Harrison
Keyword(s):  
Neutral Endopeptidase ◽  
Converting Enzyme ◽  
Simple Method ◽  
Endopeptidase 24.11 ◽  
Endothelin Converting Enzyme

Potent non-peptidic dual inhibitors of endothelin-converting enzyme and neutral endopeptidase 24.11

1997 ◽  
Vol 7 (8) ◽  
pp. 1059-1064 ◽  
Author(s):  
Stéphane De Lombaert ◽  
Lisa B. Stamford ◽  
Louis Blanchard ◽  
Jenny Tan ◽  
Denton Hoyer ◽  
...  
Keyword(s):  
Neutral Endopeptidase ◽  
Converting Enzyme ◽  
Endopeptidase 24.11 ◽  
Endothelin Converting Enzyme ◽  
Dual Inhibitors

scholarly journals Mutagenesis of Glu403 to Cys in rabbit neutral endopeptidase-24.11 (neprilysin) creates a disulphide-linked homodimer: analogy with endothelin-converting enzyme

1997 ◽  
Vol 327 (3) ◽  
pp. 925-929 ◽  
Author(s):  
V. Mien HOANG ◽  
E. Clare SANSOM ◽  
J. Anthony TURNER
Keyword(s):  
Three Dimensional ◽  
Neutral Endopeptidase ◽  
Biologically Active ◽  
Dimensional Structure ◽  
Mutant Form ◽  
Converting Enzyme ◽  
Wild Type ◽  
Endopeptidase 24.11 ◽  
Reducing Conditions ◽  
Endothelin Converting Enzyme

Neutral endopeptidase-24.11 (NEP; neprilysin; EC 3.4.24.11) and endothelin-converting enzyme (ECE) are related zinc metallopeptidases involved in the processing of biologically active peptides. Only ECE, however, exists as a disulphide-linked homodimer. The covalent linkage in rat ECE is between Cys412 in each subunit, which is equivalent to Glu403 in rabbit NEP. Here we report that directed mutagenesis of Glu403 to cysteine in rabbit NEP creates a disulphide-linked homodimer, as revealed by transient transfection in COS-1 cells and SDS/PAGE of a membrane fraction. Under reducing conditions, both the mutant (E403C) and the wild-type NEP migrate as a polypeptide of 92 kDa. However, under non-reducing conditions, the Mr of the wild type remains unchanged, whereas that of the mutant is doubled. Co-transfection of wild-type ECE and E403C NEP cDNA did not result in the production of a NEP-ECE heterodimer. Comparison of the kinetic constants for wild-type and E403C mutant NEP with either [D-Ala2,Leu5]enkephalin or 3-carboxypropanoyl-alanyl-alanyl-leucine-4-nitroanilide (Suc-Ala-Ala-Leu-NH-Np) as substrate show a decrease of approx. 50% in Vmax/Km for the mutant form. The IC50 value for inhibition of the mutant by phosphoramidon or thiorphan is increased 3-fold and 5-fold respectively. Although NEP and ECE exhibit only about 40% identity and differ substantially in substrate specificity and some other characteristics, these data indicate that they have considerable similarity in three-dimensional structure, allowing dimer formation in the mutant NEP with the disulphide link probably occurring in a hydrophilic surface loop.

ChemInform Abstract: Potent Non-Peptidic Dual Inhibitors of Endothelin-Converting Enzyme and Neutral Endopeptidase 24.11.

ChemInform ◽  
2010 ◽  
Vol 28 (36) ◽  
pp. no-no
Author(s):  
S. DE LOMBAERT ◽  
L. B. STAMFORD ◽  
L. BLANCHARD ◽  
J. TAN ◽  
D. HOYER ◽  
...  
Keyword(s):  
Neutral Endopeptidase ◽  
Converting Enzyme ◽  
Endopeptidase 24.11 ◽  
Endothelin Converting Enzyme ◽  
Dual Inhibitors
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