Hydrolysis of rat melanin-concentrating hormone by endopeptidase 24.11 (neutral endopeptidase)

Melanin-concentrating hormone (MCH) is a cyclic peptide which behaves as an antagonist of the pituitary melanotropic hormone alpha-melanocyte-stimulating hormone in fishes. Cloning of the rat MCH cDNA precursor recently revealed the presence of an additional putative peptide named NEI. The present work examined the susceptibility of these novel peptides to hydrolysis by various purified exo- and endo-peptidases including endopeptidases 24.11 (NEP), 24.15, 24.16, angiotensin-converting enzyme, leucine aminopeptidase and carboxypeptidase A. NEP attacked MCH at three sites of the molecule with an apparent affinity of about 12 microM and a kcat. of 4 min-1. The first site of cleavage was at Cys-7-Met-8, i.e. within the peptide loop formed by the internal disulphide bridge. NEP could therefore be considered as an MCH-inactivating peptidase since the degradation products generated are probably devoid of biological activity. In contrast, NEI neither inhibited the degradation of the NEP chromogenic substrate glutaryl-Phe-Ala-Phe-p-aminobenzoate nor was susceptible to proteolysis by NEP. Unlike NEP, angiotensin-converting enzyme, endopeptidase 24.15 and endopeptidase 24.16 appeared totally unable to cleave MCH, whereas the peptide was readily degraded by aminopeptidase M and carboxypeptidase A.

Download Full-text

Potent inhibition of endopeptidase 24.16 and endopeptidase 24.15 by the phosphonamide peptide N-(phenylethylphosphonyl)-Gly-l-Pro-l-aminohexanoic acid

Biochemical Journal ◽

10.1042/bj2870621 ◽

1992 ◽

Vol 287 (2) ◽

pp. 621-625 ◽

Cited By ~ 19

Author(s):

H Barelli ◽

V Dive ◽

A Yiotakis ◽

J P Vincent ◽

F Checler

Keyword(s):

Angiotensin Converting Enzyme ◽

Leucine Aminopeptidase ◽

Converting Enzyme ◽

Carboxypeptidase A ◽

Clostridium Histolyticum ◽

Endopeptidase 24.11 ◽

Potent Inhibition ◽

Hydrolysis Of ◽

Endopeptidase 24.15

A phosphonamide peptide, N-(phenylethylphosphonyl)-Gly-L-Pro-L-aminohexanoic acid, previously shown to block Clostridium histolyticum collagenases, was examined as a putative inhibitor of endopeptidase 24.16 and endopeptidase 24.15. Hydrolysis of two endopeptidase 24.16 substrates, i.e. 3-carboxy-7-methoxycoumarin (Mcc)-Pro-Leu-Gly-Pro-D-Lys-dinitrophenyl (Dnp) and neurotensin, were completely and dose-dependently inhibited by the phosphonamide inhibitor with KI values of 0.3 and 0.9 nM respectively. In addition, the phosphonamide peptide inhibited the hydrolysis of benzoyl (Bz)-Gly-Ala-Ala-Phe-(pAB) p-aminobenzoate and neurotensin by endopeptidase 24.15 with about a 10-fold lower potency (KI values of 5 and 7.5 nM respectively). The selectivity of this inhibitor towards several exo- and endo-peptidases belonging to the zinc-containing metallopeptidase family established that a 1 microM concentration of this inhibitor was unable to affect leucine aminopeptidase, carboxypeptidase A, angiotensin-converting enzyme and endopeptidase 24.11. The present paper therefore reports on the first hydrophilic highly potent endopeptidase 24.16 inhibitor and describes the most potent inhibitory agent directed towards endopeptidase 24.15 developed to date. These tools should allow one to assess the contribution of endopeptidase 24.16 and endopeptidase 24.15 to the physiological inactivation of neurotensin as well as other neuropeptides.

Download Full-text

Design and synthesis of phosphinic acids that triply inhibit endothelin converting enzyme, angiotensin converting enzyme and neutral endopeptidase 24.11

Bioorganic & Medicinal Chemistry Letters ◽

10.1016/0960-894x(96)00297-1 ◽

1996 ◽

Vol 6 (14) ◽

pp. 1629-1634 ◽

Cited By ~ 27

Author(s):

Brian A. McKittrick ◽

Andrew W. Stamford ◽

Xiaoyu Weng ◽

Ke Ma ◽

Samuel Chackalamannil ◽

...

Keyword(s):

Angiotensin Converting Enzyme ◽

Neutral Endopeptidase ◽

Converting Enzyme ◽

Design And Synthesis ◽

Endopeptidase 24.11 ◽

Endothelin Converting Enzyme ◽

Phosphinic Acids

Download Full-text

CGS 30440: A Dual Inhibitor of Angiotensin-Converting Enzyme and Neutral Endopeptidase 24.11

Cardiovascular Drug Reviews ◽

10.1111/j.1527-3466.1999.tb00002.x ◽

2006 ◽

Vol 17 (1) ◽

pp. 16-38 ◽

Cited By ~ 6

Author(s):

David S. Cohen ◽

Cynthia A. Fink ◽

Angelo J. Trapani ◽

Randy L. Webb ◽

Patricia A. Zane ◽

...

Keyword(s):

Angiotensin Converting Enzyme ◽

Neutral Endopeptidase ◽

Converting Enzyme ◽

Dual Inhibitor ◽

Endopeptidase 24.11

Download Full-text

Dual inhibition of angiotensin converting enzyme and neutral endopeptidase by omapatrilat in rat in vivo

Pharmacological Research ◽

10.1006/phrs.2001.0875 ◽

2001 ◽

Vol 44 (5) ◽

pp. 411-418 ◽

Cited By ~ 20

Author(s):

Tom Bäcklund ◽

Eeva Palojoki ◽

Tina Grönholm ◽

Anders Eriksson ◽

Olli Vuolteenaho ◽

...

Keyword(s):

Angiotensin Converting Enzyme ◽

Neutral Endopeptidase ◽

Converting Enzyme ◽

Dual Inhibition

Download Full-text

Novel activity of angiotensin-converting enzyme. Hydrolysis of cholecystokinin and gastrin analogues with release of the amidated C-terminal dipeptide

Biochemical Journal ◽

10.1042/bj2620125 ◽

1989 ◽

Vol 262 (1) ◽

pp. 125-130 ◽

Cited By ~ 39

Author(s):

P Dubreuil ◽

P Fulcrand ◽

M Rodriguez ◽

H Fulcrand ◽

J Laur ◽

...

Keyword(s):

Angiotensin Converting Enzyme ◽

Chloride Ion ◽

Peptide Bond ◽

Major Product ◽

Enzyme Hydrolysis ◽

Ion Concentration ◽

Converting Enzyme ◽

Rabbit Lung ◽

Hydrolysis Of

ACE (angiotensin-converting enzyme; peptidyl dipeptidase A; EC 3.4.15.1), cleaves C-terminal dipeptides from active peptides containing a free C-terminus. We investigated the hydrolysis of cholecystokinin-8 [CCK-8; Asp-Tyr(SO3H)-Met-Gly-Trp-Met-Asp-Phe-NH2] and of various gastrin analogues by purified rabbit lung ACE. Although these peptides are amidated at their C-terminal end, they were metabolized by ACE to several peptide fragments. These fragments were analysed by h.p.l.c., isolated and identified by comparison with synthetic fragments, and by amino acid analysis. The initial and major site of hydrolysis was the penultimate peptide bond, which generated a major product, the C-terminal amidated dipeptide Asp-Phe-NH2. As a secondary cleavage, ACE subsequently released di- or tri-peptides from the C-terminal end of the remaining N-terminal fragments. The cleavage of CCK-8 and gastrin analogues was inhibited by ACE inhibitors (Captopril and EDTA), but not by other enzyme inhibitors (phosphoramidon, thiorphan, bestatin etc.). Hydrolysis of [Leu15]gastrin-(14-17)-peptide [Boc (t-butoxycarbonyl)-Trp-Leu-Asp-Phe-NH2] in the presence of ACE was found to be dependent on the chloride-ion concentration. Km values for the hydrolysis of CCK-8, [Leu15]gastrin-(11-17)-peptide and Boc-[Leu15]gastrin-(14-17)-peptide at an NaCl concentration of 300 mM were respectively 115, 420 and 3280 microM, and the catalytic constants were about 33, 115 and 885 min-1. The kcat/Km for the reactions at 37 degrees C was approx. 0.28 microM-1.min-1, which is approx. 35 times less than that reported for the cleavage of angiotensin I. These results suggest that ACE might be involved in the metabolism in vivo of CCK and gastrin short fragments.

Download Full-text

Endothelial angiotensin-converting enzyme and neutral endopeptidase in isolated human umbilical vein: An effective bradykinin inactivation pathway

European Journal of Pharmacology ◽

10.1016/j.ejphar.2011.05.045 ◽

2011 ◽

Vol 667 (1-3) ◽

pp. 271-277 ◽

Cited By ~ 6

Author(s):

Wanda Nowak ◽

Andrea Emilse Errasti ◽

Arnaldo Raúl Armesto ◽

Natalia Lucía Santín Velazque ◽

Rodolfo Pedro Rothlin

Keyword(s):

Angiotensin Converting Enzyme ◽

Umbilical Vein ◽

Neutral Endopeptidase ◽

Converting Enzyme ◽

Human Umbilical Vein ◽

Bradykinin Inactivation

Download Full-text

Kinetic probes for inter-domain co-operation in human somatic angiotensin-converting enzyme

Biochemical Journal ◽

10.1042/bj20050702 ◽

2005 ◽

Vol 391 (3) ◽

pp. 641-647 ◽

Cited By ~ 16

Author(s):

Olga E. Skirgello ◽

Peter V. Binevski ◽

Vladimir F. Pozdnev ◽

Olga A. Kost

Keyword(s):

Angiotensin Converting Enzyme ◽

Enzymatic Activity ◽

Active Site ◽

The Other ◽

Converting Enzyme ◽

Human Enzyme ◽

Independent Functions ◽

The Common ◽

The Individual ◽

Hydrolysis Of

s-ACE (the somatic form of angiotensin-converting enzyme) consists of two homologous domains (N- and C-domains), each bearing a catalytic site. Negative co-operativity between the two domains has been demonstrated for cow and pig ACEs. However, for the human enzyme there are conflicting reports in the literature: some suggest possible negative co-operativity between the domains, whereas others indicate independent functions of the domains within s-ACE. We demonstrate here that a 1:1 stoichiometry for the binding of the common ACE inhibitors, captopril and lisinopril, to human s-ACE is enough to abolish enzymatic activity towards FA {N-[3-(2-furyl)acryloyl]}-Phe-GlyGly, Cbz (benzyloxycarbonyl)-Phe-His-Leu or Hip (N-benzoylglycyl)-His-Leu. The kinetic parameters for the hydrolysis of seven tripeptide substrates by human s-ACE appeared to represent average values for parameters obtained for the individual N- and C-domains. Kinetic analysis of the simultaneous hydrolysis of two substrates, Hip-His-Leu (S1) and Cbz-Phe-His-Leu (S2), with a common product (His-Leu) by s-ACE at different values for the ratio of the initial concentrations of these substrates (i.e. σ=[S2]0/[S1]0) demonstrated competition of these substrates for binding to the s-ACE molecule, i.e. binding of a substrate at one active site makes the other site unavailable for either the same or a different substrate. Thus the two domains within human s-ACE exhibit strong negative co-operativity upon binding of common inhibitors and in the hydrolysis reactions of tripeptide substrates.

Download Full-text