Characterisation of neutral endopeptidase 3.4.24.11 (NEP) in the kidney: comparison between normotensive, genetically hypertensive and experimentally hypertensive rats.
Neutral endopeptidase 3.4.24.11 (NEP) has been identified as the major atrial natriuretic factor (ANF) degrading enzyme in rat kidney, therefore, suggesting a possible role for this enzyme in blood volume and pressure regulation. Various experimentally induced and genetically hypertensive rat models have been used to test NEP inhibitors. The presence of different isoforms of NEP in the various hypertensive rat models would have relevance when searching for novel NEP inhibitors. Therefore, we compared the properties of NEP in kidney cortex homogenates in order to test for possible differences in the following hypertensive rat models and their appropriate controls: spontaneously hypertensive rats (SHR), Wistar Kyoto strain (WKY), DOCA-salt hypertensive rats, and Sprague Dawley control rats (SD). No relevant differences were found when comparing the following parameters: (1) specific activity (mean: 204 U/mg protein), (2) Michaelis constant (mean: 280 microM), (3) IC50 of thiorphan (mean: 6.5 nM) and phosphoramidon (mean: 54 nM), (4) pH profiles (optimum at pH 8.0), (5) heat inactivation profiles (half-life 20 min at 65 degrees C), (6) immunotitration of kidney cortex homogenates, (7) molecular weight as determined by gel filtration (92,000 Dalton) and (8) affinity chromatography with concanavalin A. Without evidence for the presence of different NEP isoforms, it is unlikely that divergent findings in DOCA-salt rats and SHR using a given NEP inhibitor are due to isoforms of NEP.
Schulz
R
R
Research Department, CIBA-GEIGY Corporation, Summit, New Jersey 07901.
Sakane
Y
Y
Berry
C
C
Ghai
R
R
D000818:Animals; D001835:Body Weight; D004247:DNA / Q000032:analysis; D003900:Desoxycorticosterone / Q000494:pharmacology; D006020:Glycopeptides / Q000494:pharmacology; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D006973:Hypertension / Q000139:chemically induced / Q000201:enzymology* / Q000235:genetics; D007672:Kidney Cortex / Q000201:enzymology*; D015260:Neprilysin / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D009929:Organ Size; D011489:Protein Denaturation; D011506:Proteins / Q000032:analysis; D051381:Rats; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY; D017207:Rats, Sprague-Dawley; D012492:Salts / Q000494:pharmacology; D015244:Thiorphan / Q000494:pharmacology
D006020:Glycopeptides; D011506:Proteins; D012492:Salts; D003900:Desoxycorticosterone; D004247:DNA; D015244:Thiorphan; D015260:Neprilysin; C008890:phosphoramidon