Biochemical and immunological properties of urinary angiotensinase A and dipeptidylaminopeptidase IV. Their use as markers in patients with renal cell injury.
Dipeptidyl peptidase IV (EC 3.4.14.5) and angiotensinase A (EC 4.4.11.7) were purified to homogeneity from pooled urine concentrate of patients with renal damage, using ultrafiltration, ammonium sulphate precipitation, lectin affinity chromatography, FPLC-ion-exchange(Mono-Q-)chromatography, and FPLC-gel filtration (Superdex). Based on the specific enzyme activity of the starting material, dipeptidyl peptidase IV was enriched 1629 fold, angiotensinase A 1183 fold. The relative molecular masses, Michaelis constants and isoelectric points were determined. Negative staining of the purified enzymes revealed globular proteins (5-7 nm). Antisera raised against dipeptidyl peptidase IV and angiotensinase A reacted specifically with tubular and, in the case of anti-angiotensinase A sera, with tubular and glomerular structures. In addition, urinary membrane vesicles of proximal tubule origin were eluted with the void volume (Superdex-gel filtration), indicating heavy epithelial cell disintegration. Both soluble tissue enzymes (dipeptidyl peptidase IV, angiotensinase A) and vacuolar blebs shed from epithelia contribute to proteinuria, as was shown in patients with glomerulonephritis, interstitial nephritis, diabetic nephropathy and, for angiotensinase A, in patients with essential arterial hypertension.
Klinikum der J. W. Goethe-Universität, Abteilung für Nephrologie, Zentrum Innere Medizin, Frankfurt am Main.
J E
JE
Scherberich
J
J
Wiemer
W
W
Schoeppe
D000293:Adolescent; D000328:Adult; D000626:Aminopeptidases / Q000276:immunology / Q000302:isolation & purification / Q000652:urine*; D000818:Animals; D000917:Antibody Formation; D000941:Antigens / Q000276:immunology; D015415:Biomarkers / Q000652:urine; D018819:Dipeptidyl Peptidase 4; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000276:immunology / Q000302:isolation & purification / Q000652:urine*; D005260:Female; D043384:Glutamyl Aminopeptidase; D006801:Humans; D007674:Kidney Diseases / Q000201:enzymology* / Q000652:urine; D007684:Kidney Tubules / Q000201:enzymology / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000302:isolation & purification / Q000652:urine; D008566:Membranes / Q000201:enzymology; D008875:Middle Aged; D011507:Proteinuria / Q000652:urine; D011817:Rabbits
D000941:Antigens; D015415:Biomarkers; D008565:Membrane Proteins; D000626:Aminopeptidases; D043384:Glutamyl Aminopeptidase; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D018819:Dipeptidyl Peptidase 4