Degradation and inactivation of human atrial natriuretic peptide by human pulmonary plasma membranes.
Atrial natriuretic peptide (ANP) is extracted from plasma during its passage through the lungs. ANP is metabolized in rat lung membrane preparations by the enzyme neutral endopeptidase-24.11 (EC 3.4.24.11), but the hydrolysis of ANP in human lung has not been characterized. In the present study synthetic human atrial natriuretic peptide 1-28 (alpha-hANP) was separately incubated with human pulmonary plasma membranes from two non-smoking patients, and the major degradation products were separated from alpha-hANP by reverse-phase high pressure liquid chromatography. The degradation products were identified by sequence analysis and by mass-spectrometry, and biological activity was studied in vitro by exposing precontracted rabbit pulmonary arteries to alpha-hANP and the degradation products. The initial cleavage appeared, with membrane preparations from both patients, in the central ring structure between Arg14 and Ile15, followed by a cleavage of the bond Arg3-Arg4 at the N-terminal region of the peptide. The biological activity of this ring-opened product was about 1/500 of the activity of uncleaved alpha-hANP. Cleavage of the Arg3-Arg4 or Arg14-Ile15 bonds could not be inhibited by EDTA, iodoacetamide, benzamidine hydrochloride or pepstatin A. Neither did phosphoramidon (1 microM) or thiorphan (1 microM) inhibit the hydrolysis, indicating the presence in human lung of an ANP-degrading enzyme different from endopeptidase-24.11.
Lindberg
B F
BF
Department of Clinical Pharmacology, University Hospital of Lund, Sweden.
Andersson
K E
KE
D000368:Aged; D000595:Amino Acid Sequence; D000818:Animals; D009320:Atrial Natriuretic Factor / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006801:Humans; D007700:Kinetics; D008168:Lung / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D008969:Molecular Sequence Data; D015260:Neprilysin / Q000037:antagonists & inhibitors / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D011817:Rabbits
D011480:Protease Inhibitors; D009320:Atrial Natriuretic Factor; D015260:Neprilysin