Processing of angiotensin peptides by NG108-15 neuroblastoma x glioma hybrid cell line.
The metabolism of angiotensin (Ang) peptides was studied in NG108-15 neuroblastoma x glioma hybrid cells which express Ang II receptors, renin, dipeptidyl carboxypeptidase A (converting enzyme), as well as Ang I and Ang II. In these experiments, 0.2 nM of either 125I-Ang I or 125I-Ang II was incubated with intact cell monolayers and the medium was analyzed for 125I-products by high performance liquid chromatography. The major product generated from the metabolism of labeled Ang I or Ang II was identified as the amino-terminal heptapeptide Ang-(1-7). N-benzyloxycarbonyl-prolyl-prolinal (ZPP), a specific inhibitor of prolyl endopeptidase, inhibited the formation of Ang-(1-7) from Ang I by 35%. Complete inhibition of Ang-(1-7) generation was attained with p-chloromercuriphenyl-sulfonate, which suggests that a sulfhydryl-containing peptidase other than prolyl endopeptidase is also involved in Ang-(1-7) formation. Ang II was observed to be a minor product resulting from Ang I metabolism. Although the converting enzyme inhibitor enalaprilat (MK-422) significantly reduced Ang II formation, it had no effect on the levels of Ang-(1-7). These findings demonstrate a preferential processing of Ang I into Ang-(1-7) which is not dependent on the prior formation of Ang II.
Chappell
M C
MC
Department of Brain, Cleveland Clinic Foundation, OH 44195-5070.
Tallant
E A
EA
Brosnihan
K B
KB
Ferrario
C M
CM
D002731:4-Chloromercuribenzenesulfonate / Q000494:pharmacology; D000704:Analysis of Variance; D000803:Angiotensin I / Q000378:metabolism*; D000804:Angiotensin II / Q000096:biosynthesis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D004151:Dipeptides / Q000494:pharmacology; D015773:Enalaprilat / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D006822:Hybrid Cells; D010446:Peptide Fragments / Q000096:biosynthesis; D010455:Peptides / Q000032:analysis*; D000085822:Prolyl Oligopeptidases; D012697:Serine Endopeptidases; D014407:Tumor Cells, Cultured
D004151:Dipeptides; D010446:Peptide Fragments; D010455:Peptides; D000804:Angiotensin II; D002731:4-Chloromercuribenzenesulfonate; C038541:N-benzyloxycarbonylprolylprolinal; D000803:Angiotensin I; D010450:Endopeptidases; D012697:Serine Endopeptidases; D000085822:Prolyl Oligopeptidases; D015773:Enalaprilat; C118790:angiotensin I (1-7)