A reliable radiometric assay for the determination of angiotensin I-converting enzyme activity in urine.
We present a radiometric assay for the determination of urinary angiotensin-converting enzyme activity, using benzoyl-[1-14C]glycyl-L-histidyl-L-leucine as the substrate. An optimal pH of 8.3, an optimal chloride concentration of 0.375 mol/l and complete inhibition by EDTA-Na2, captopril and enalaprilat confirm the specificity of the assay. Comparison of dialysis and ultrafiltration for concentration of urine showed the existence of angiotensin-converting enzyme inhibitors in human urine. Dialysis against water was the more effective method for avoiding enzyme inhibition. After dialysis of urine, the assay was linear with time and with enzyme concentration; it was highly sensitive (60 mU/l) and showed good reproducibility. Under our technical conditions, we found angiotensin-converting enzyme activity in urine samples with quantitatively abnormal protein contents, but not in normal urine. Urinary angiotensin-converting enzyme did not correlate with proteinuria nor with water-salt parameters or creatinine. We confirm the kidney tubular epithelial origin of the enzyme, and propose the use of our assay to study urinary angiotensin-converting enzyme as a marker of renal tubular damage.
Baudin
B
B
Laboratoire de Biochimie A, Hôpital Saint-Antoine, Paris, France.
Bénéteau-Burnat
B
B
Baumann
F C
FC
Giboudeau
J
J
D000293:Adolescent; D000328:Adult; D000803:Angiotensin I / Q000652:urine; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000652:urine; D003956:Dialysis; D004492:Edetic Acid / Q000494:pharmacology; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008297:Male; D007703:Peptidyl-Dipeptidase A / Q000652:urine*; D011874:Radiometry / Q000379:methods* / Q000592:standards
D000806:Angiotensin-Converting Enzyme Inhibitors; D000803:Angiotensin I; D004492:Edetic Acid; D007703:Peptidyl-Dipeptidase A