Please use this identifier to cite or link to this item: http://ahro.austin.org.au/austinjspui/handle/1/9799
Title: Localization and characterization of neutral metalloendopeptidase (EC 3.4.24.11), the degradative enzyme for atrial natriuretic peptide, in rat kidney using a radioiodinated neutral metalloendopeptidase inhibitor.
Authors: Kanazawa, M;Casley, David J;Sybertz, E J;Haslanger, M F;Johnston, Colin I
Affiliation: Department of Medicine, University of Melbourne, Austin Hospital, Heidelberg, Victoria, Australia.
Issue Date: 1-Jun-1992
Citation: The Journal of Pharmacology and Experimental Therapeutics; 261(3): 1231-7
Abstract: Atrial natriuretic peptide (ANP) is rapidly degraded by neutral metalloendopeptidase (EC 3.4.24.11, NEP), with the kidney being a major site of ANP clearance. NEP has been anatomically localized in the rat kidney by in vitro autoradiography and the active site studied by a radioinhibitor binding assay (RIBA) using a newly developed radioinhibitor as a radioligand. SCH47896 is a phenolic derivative of SCH39370, a potent specific inhibitor of NEP, which can be radioiodinated with 125I. NEP catalytic activity in the rat kidney was inhibited by SCH47896 and its di-iodo analog SCH48446. Specific binding of [125I]SCH47896 to renal plasma membranes fitted a single-site model with Kd = 43.3 nM and maximal binding site density = 13.8 pmol/mg protein. Thus, [125I]SCH47896 retains full enzymatic inhibitory activity and full binding to the active site of the NEP. Autoradiographs using [125I]SCH47896 demonstrated maximal binding to deep proximal renal tubules. This binding was displaced in a dose-dependent manner by NEP inhibitors. Renal NEP was inhibited by SCH39370. Inhibition of ANP degradation by NEP in the kidney by the new NEP or atriopeptidase inhibitors may explain their natriuretic and diuretic effect in the absence of changes in plasma ANP levels. These studies will allow investigation of the regulation of NEP and the role inhibition of tissue NEP plays in the actions of the new atriopeptidase inhibitors. Furthermore, this method of radioinhibitor binding is applicable to any enzyme, provided a suitable radioligand can be constructed.
Internal ID Number: 1534841
URI: http://ahro.austin.org.au/austinjspui/handle/1/9799
URL: http://www.ncbi.nlm.nih.gov/pubmed/1534841
Type: Journal Article
Subjects: Animals
Atrial Natriuretic Factor.metabolism
Autoradiography
Binding, Competitive
Cell Membrane.drug effects.metabolism
Dipeptides.pharmacology
Drug Interactions
Glycopeptides.pharmacology
Iodine Radioisotopes
Kidney.drug effects.enzymology
Male
Neprilysin.antagonists & inhibitors.metabolism
Phenols.metabolism.pharmacology
Phenylalanine.analogs & derivatives.metabolism.pharmacology
Rats
Rats, Inbred Strains
Thiorphan.pharmacology
Appears in Collections:Journal articles

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