title: Isolation and partial characterization of angiotensinase A and aminopeptidase M from urine and human kidney by lectin affinity chromatography and high-performance liquid chromatography.
abstract: Angiotensinase A (ATA) and aminopeptidase M (APM) were partially purified from human urine specimens and human kidney particles using wheat germ lectin affinity chromatography, anion-exchange Fast Protein Liquid Chromatography (FPLC) (Mono Q), chromatofocusing (Mono P, FPLC) and Superose 12 gel filtration. APM, a globular 5-nm glycoprotein, is localized in the brush border membrane of the proximal tubule; angiotensin II-degrading ATA is present on glomerular endothelia and podocytes and, to lesser extent, in the brush border. For the first time, both peaks of ATA and APM activity from urine samples were separated by the above-mentioned techniques with only slight overlap; ATP (146,000 dalton: pI4.8) was enriched more than 20-fold and APM (153,000 dalton, pI4.7) more than 50-fold compared with the activity of the starting material. Using similar separation steps, ATA and APM solubilized from kidney particles could not be resolved into two distinct peak fractions, however, except after hydrophobic interaction chromatography. Thus urine is a major source for the preparation of individual ATA and APM fractions, necessary to generate specific anti-enzyme antibodies for diagnostic purposes.
authors.affiliation: Department of Nephrology, Centre of Internal Medicine, J. W. Goethe University, Frankfurt am, Main, F.R.G.
authors.forename: J E
authors.initials: JE
authors.lastname: Scherberich
authors.forename: J
authors.initials: J
authors.lastname: Wiemer
authors.forename: C
authors.initials: C
authors.lastname: Herzig
authors.forename: P
authors.initials: P
authors.lastname: Fischer
authors.forename: W
authors.initials: W
authors.lastname: Schoeppe
mesh_terms: D000626:Aminopeptidases / Q000302:isolation & purification* / Q000652:urine; D018826:CD13 Antigens; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D043384:Glutamyl Aminopeptidase; D006801:Humans; D007525:Isoelectric Focusing; D007668:Kidney / Q000201:enzymology*; D014909:Wheat Germ Agglutinins
chemical_list: D014909:Wheat Germ Agglutinins; D000626:Aminopeptidases; D018826:CD13 Antigens; D043384:Glutamyl Aminopeptidase